Conversations with Laureates
Born November 22, 1912 in Cleveland, Ohio
Married Mary Connor in October, 1936
Education
A.B. Dartmouth College
M.D. Harvard Medical School
Chronology of Employment
1936 - 1937 Research Fellow in Medicine, Harvard Medical School
1937 - 1938 Research Fellow in Medicine, Harvard Medical School Faculty
1938 - 1939 Intern in Medicine, University Hospitals, Cleveland
1939 - 1940 Moseley Traveling Fellow of Harvard, and Visiting Investigator, Carlsberg Laboratory, Copenhagen (Kai Linderstrom Lang)
1940 - 1941 Assistant Physician, Huntington Memorial Hospital, Boston
1941 - 1942 Finney Howell Research Fellow, Rockefeller Institute for Medical Research (Max Bergman)
1942 - 1956 Instructor to Assoc. Prof. Medicine, Harvard Medical School
1947 - 1946 Tutor in Biochemical Sciences, Harvard University
1952 Visiting Fellow, Department of Chemistry (Linus Paling), California Institute of Technology
1950 - 1956 Associate Physician, Massachusetts General Hospital
1954 - 1956 Chairman, Committee on Research, Massachusetts General Hospital
1955 - 1979 Collis P. Huntington Professor of Oncologic Medicine and Director, John Collins Warren Laboratories of the Huntington Memorial Hospital of Harvard University
1955 - 1979 Physician, Massachusetts General Hospital
1954 - 1962 Commonwealth Fellow, University Chemical Laboratory, Cambridge University, England (Lord Todd)
1964 - 1965 President, American Association for Cancer Research
1956 - 1971 Chairman, Executive Committee, Departments of Medicine, Harvard Medical School
1975, 1978 Fogarty Scholar, National Cancer Institute, Bethesda, Maryland
1979 - 1997 Principal Investigator, Worcester Foundation for Biomedical Research, Shrewsbury, Massachusetts
1983 to present Senior Physician, Massachusetts General Hospital
1984 to present Honorary Physician, Massachusetts General Hospital
1991 - 1997 Chairman, Scientific Advisory Board, Hybridon, Inc.
1998 to present Chairman, Scientific Advisory Board, Hybridon, Inc.
1998 to present
Senior Scientist, Massachusetts General Hospital
Important Places
American Association for Cancer Research
The AACR facilitates research among varied scientific disciplines, manages four cancer-related journals, and supports cancer research through fellowships and programs. Links to AACR programs and information are available on their Web site.
Carlsberg Research Center (Carlsberg Laboratories)
The Carlsberg Research Center, incorporating Carlsberg Laboratories as well as the Research Laboratory for the brewery, and Process and Product development, focuses on sciences related to brewing. The center offers information on its departments of chemistry, physiology and yeast genetics, seminars, meetings and related links.
The Center for Molecular Biology of RNA
The Center, a part of the University of California at Santa Cruz and based at the Sinsheimer Labs, offers general information on RNA, related links, images, and faculty publications.
Harvard Medical School
The Harvard Medical School Web site offers information on faculty and staff research, HMS publications, links to affiliated institutions and hospitals, and a section for education and admissions.
Howard Hughes Medical Institute (HHMI) Biopolymer/Keck Foundation Biotechnology Resource Laboratory
The Keck Foundation, a facility of the Yale School of Medicine, researches areas of nucleic acid chemistry, peptide synthesis, and protein chemistry. The site includes links to Yale Microarray Database.
IMB Jena Image Library of Biological Macromolecules
The IMB Jena Image Library provides 3-dimensional biopolymer structures of all entries in the Protein Data Bank and the Nucleic Acid Database. Basic architectural information on the biopolymer structures is also available.
Massachusetts General Hospital
Massachusetts General Hospital offers information for patients and visitors, on departments and programs, and links to research resources, including the Treadwell Library.
Massachusetts Institute of Technology
MIT's Web site includes links to news, events, university research and resources as well as academic information.
National Institute of Health
The NIH has a long history of scientific research, with specific federal attention on medical knowledge and advances for the public. Information on NIH programs and institutes, resources, news and events are all available on their Web site.
The Nucleic Acid Database
The Nucleic Acid Database, maintained by Rutgers University, "assembles and distributes structural information about nucleic acids." The site has access to the Nucleic Acid Database (NDB), the NMR-Nucleic Acid Database and the DNA-Binding Protein Database, along with links to crystal structure data, database archives, and atlases (using text and graphics).
The RCSB Protein Data Bank
The Protein Data Bank, a part of the Research Collaboratory for Structural Bioinformatics, is a resource for macromolecular structures. The site also offers educational information and press releases as well as news, file formats, dictionaries and links to related information.
The Ribosomal Database Project
The Ribosomal Database Project (RDP), previously a part of the University of Illinois at Urbana-Champagne and now supported by Michigan State University, offers online data analyses of RNA sequences with phylogenic information and annotated rRNA sequences.
The RNA Society
The RNA Society facilitates the sharing of information regarding experimental research in the fields of biochemistry, molecular, evolutionary and structural biology, biomedicine, genetics, virology and chemistry. The Society Web site includes information on the 2000 Conference, news and events, related links, and The RNA Journal.
The RNA Structure Database
RNABase compiles raw structural information from the Nucleic Acid Database (NDB) and the Protein Data Bank (PDB) in an annotated database with information on RNA structure and nomenclature, database analysis, RNABase Bibliography and online resources.
The Rockefeller University
Originally the Rockefeller Institute of Medical Research, The Rockefeller University offers information on its research projects, links to their hospital, and information on clinical trials, featured news and academic programs.
Honors
1932-1934 - Phi Beta Kappa, Magna Cum Laude, Highest Distinction in Anatomy
1946 - John Collins Warren Triennial Prize (with Dr. Fritz Lipmann)
1947 - John Collins Warren Triennial Prize
1948 - Harvey Society Lecture
1962 - First Jubilee Lectureship, Biochemical Society, London
1963 - James Ewing Award
1965 - Borden Award in Medical Sciences
1966 - D.Sc., University of Utrecht (hon.)
1968 - American Cancer Society National Award
1970 - Passano Award 1971 D.Sc., Columbia University (hon.)
1982 - D.Sc., Harvard University (hon.)
1983 - D.Sc., Roger Williams College (hon.)
1984 - D.Sc., Dartmouth College (hon.)
1991 - Presidential Medal of Science
1992 - Hudson Hoagland Award
1994 - D.Sc., U. Massachusetts (hon.)
1995 - City of Medicine Award, Durham, NC
1996 - Century 2000 Science Award, City of Worcester, MA
1996 - Lasker Lifetime Achievement Award
1997 - American Society of Biological Chemistry and Molecular Biology Merck Award
1998 - Annual Orator, Mass. Medical Society
1999 - John Collins Warren Triennial Prize (with Dr. Christiane Nüsslein-Volhard)
Societies
American Academy of Arts and Sciences
American Society of Biological Chemistry
American Association for Cancer Research
Association of American Physicians
National Academy of Sciences
Danish Academy of Sciences and Letters (hon.)
National Academy of Medicine
Published Work
1. Zamecnik, P.C. and Koletsky, S.: Lack of carcinogenic potency of sulfanilamide and prontosil soluble in mice. Proc. Soc. Exp. Biol. And Med. 42: 391-392, 1939.
2. Zamecnik, P.C.: Studies on enzymatic histochemistry. XXXV. Application of the Cartesian Diver technique to measurements of the respiration of cells grown in tissue culture. Comp. Rendus des Travaux du Laboratoire Carlsberg, Serie Chimique 24: 59-67, 1941.
3. Gusberg, S.B., Zamecnik. P.C. and Aub, J.C.: The distribution of injected organic diselenides in tissue of tumor-bearing animals. J. Pharm. Exp. Therapy 71: 239-245, 1941.
4. Zamecnik, P.C., Lavin, G.I. and Bergmann, M.: Microphotometric determination of the rate of enzymatic proteolysis. J. Biol. Chem. 158: 537-545, 1945.
5.Aub, JC. Brues, A.M., Kety, S.S. Nathanson, I.T., Pope, A. and Zamecnik, P.C.: Bacteria and the toxic factor in shock. War Medicine 5: 71-73, 1944.
6. Zamecnik, P.C., Stephenson, M.L. and Cope, O,: Peptidase activity of lymph and serum after burns. J Biol. Chem. 158: 135-144, 1945.
7. Adamson, J.D., Jolliffe, M., Kruse, H.D., Lowry, O.H., Moore, P.E., Platt, B.S., Sebrell, W.H., Tice, J.W., Tisdall, F.F., Wilder, R.M. and Zamecnik, P.C.: Medical survey of nutrition in Newfoundland. Can. Med. Assoc. J. 52: 227-250, 1946.
8. Zamecnik, P.D. and Stephenson. M.L.: Distribution of catheptic enzymes in the hog kidney. J. Biol. Chem. 159: 625-629, 1945.
9. Nathanson, I.T., Nutt, A.L., Pope, A., Zamecnik, P.C., Aub, J.C, Bures A.M. and Kety, S.S.: The toxic factors in experimental traumatic shock. I. Physiological effects of muscle ligation in the dog. J. Clin. Invest. 24: 829-834, 1945.
10.Brues, A.M., Cohn, W.E., Kety, S.S. Nathanson, I.T., Nutt, A.L., Tibbetts, D.M., Zamecnik, P.C. and Aub, ,J.C.: The toxic factors in experimental traumatic shock. II. Studies on electrolyte and water balance in shock. J. Clin. /nvest. 24: 835-838, 1945.
11.Kety, S.S., Nathanson I.T., Nutt, A.L, Pope, A., Zamecnik, P.C., Aub. J.C. and Brues, A.M.: The toxic factors in experimental traumatic shock, III. Shock accompanying muscle ischemia and loss of vascular fluid. J. C/in. Invest. 24: 839-844, 1945.
12. Aub, J.C., Brues, A.M., Kety, S.S., Nathanson, I.T., Nutt, A.L., Pope, A. and Zamecnik, P.C.: The toxic factors in experimental traumatic shock. IV. The effects of intravenous injections of the effusion from ischemic muscle. .J Clin. Invest 24: 845-849, 1945.
13. Zamecnik, P.C., Aub, J.C., Brues, A.M., Kety, S.S., Nathanson, I.T., Nutt, A.L. and Pope, A.: The toxic factors in experimental traumatic shock. V. Chemical and enzymatic properties of muscle exudate. J. Clin. Invest. 24: 850-855, 1945.
14. Pope, A., Zamecnik, P.C., Aub, J.C, Brues, A.M., Dubos, R.J., Nathanson, I.T., and Nutt, A.L.: The toxic factors in experimental traumatic shock. VI. The toxic influence of the bacterial flora, particularly Clostridium welchii in exudates of ischemic muscle. J C/in. Invest. 24: 856-863, 1945.
15. Zamecnik, P.D., Folch, J., and Brewster, L: Protection of animals against C. welchii (Type A) by injection of certain purified lipids. Proc. Soc.Exp.Biol. Med. 60: 30-39, 1945.
16 Zamecnik, P.C., Brewster, L.E. and Lipmann, F.: A manometric method for measuring the activity of the C. welchii lecithinase and a description of certain properties of this enzyme. J Exp. Med. 85:381-394, 1947.
17. Zamecnik, P.C. and Lipmann, F.: A study of the competition of lecithin and antitoxin for C. welchii lecithinase. J. Exp. Med. 85: 395-403, 1947.
18. Zamecnik, P.C., Nathanson, I.T., and Aub, J.C.: The toxic factors in experimental traumatic shock. VIII. Physiologic action of C. welchii (Type A) toxins in dogs. J Clin. Invest. 26: 394-403, 1947.
19. Aub, J.C., Zamecnik, P.C. and Nathanson, I.T.: The toxic factors in experimental traumatic shock. IX. Physiologic action of C. oedematiens (Novyi) toxin in dogs. J Clin. Invest. 26: 404-410, 1947.
20. Krayer, O., Aub, J.C., Nathanson, I.T. and Zamecnik, P.C.: The toxic factors in experimental traumatic shock. Z. The influence of antitoxin upon the action of C. oedematiens toxin in the heart-lung preparation of the dog. J Clin. Invest. 26: 411-415, 1947.
21. Zamecnik, P.C. and Stephenson, M.L.: A manometric method for determining the kinetics of an enzymatic hydrolysis of peptides. J. Biol .Chem. 160: 349-357, 1947.
22. Morrison, L.R., and Zamecnik, P.C.: Experimental demyelination by means of enzymes, especially in a toxin of Clostridium welchii. Arch. Neurol. and Psych. 63: 367-381,1950.
23. Zamecnik, P.C. and Stephenson M.L.: Activity of catheptic enzymes in p-dimethylamino-azobenzene-induced hepatomas. Cancer Research 7: 326-332, 1947.
24. Frantz, I.D., Jr., Zamecnik, P.C., Reese, J.W. and Stephenson. M.L.: The effect of dinitrophenol on the incorporation of alanine labeled with radioactive carbon into the proteins of slices of normal and malignant rat liver. J. Biol. Chem. 174: 773-774, 1948.
25. Zamecnik, P.C., Frantz, I.D., Jr., Loftfield, RB. and Stephenson, M.L.: Incorporation in vitro of radioactive carbon from carboxyl-labeled DL-alanine and glycine into proteins of normal and malignant rat livers. J. Biol. Chem. 174: 773-774, 1948.
26. Zamecnik, P.C. and Stephenson, M.D.: A comparison of activators of proteolytic enzymes and peptidases in normal rat livers and hepatomas. Cancer Research 9: 3-1 1, 1949.
27. Zamecnik, P.C., Loftfield, R.B., Stephenson, M.L. and Williams, C.M.: Biological synthesis of radioactive silk. Science 109: 624-626, 1949.
28. Aykroyd, W.R., Jolliffe, N., Moore, P,E., Sebrell, W.H, Shank, RE., Tisdall, F.F,, Wilder, R.M. and Zamecnik, P.C.: Medical survey of nutrition in Newfoundland, 1948. Can. Med. Assoc. J. 60: April, 1949.
29. Zamecnik, P.C. and Aub, J.C.: Growth. Ann Review of Physiology, 1950, pp 71-100.
30. Zamecnik, P.C. and Frantz, I.D., Jr.: Peptide bond synthesis in normal and malignant tissues. Cold Spring Harbor Symp. Quant. Bio/. 14: 199-208, 1950.
31. Frantz, I.D., Jr. and Zamecnik, P.C.: Use C14-labeled amino acids in the study of peptide bond synthesis. Symposium on Nutrition. Vol. II. Plasma Proteins, C.C. Thomas. Pub., 1950, pp 85-107.
32. Zamecnik, P.C.: The use of labeled amino acids in the study of protein metabolism of normal and malignant tissues: A review. Cancer Research 10: 659-667, 1950.
33. Emerson, W.J., Zamecnik, P.C. and Nathanson, I.T.: The effect of sex hormones on hepatic and renal lesions induced in rats by a choline-deficient diet. Endocrinology 48: 545-559, 1951.
34 Zamecnik, P.C., Loftfield, R,B., Stephenson, M.L. and Steele, J.M.: Studies on the carbohydrate and protein metabolism of the rat hepatoma. Cancer research 11: 592-602, 1951.
35. Zamecnik, P.C.: Biochemistry of neoplastic tissue. Ann. Rev. Biochem. 21:411-430, 1952.
36. Siekevitz, P. and Zamecnik, P.C.: In vitro incorporation of l-C14-DL-alanine into proteins of rat liver granular fractions. Fed. Proc. 10: 265, 1951 (Abstract).
37. Zamecnik, P.C.: Studies of relationships of protein and carbohydrate metabolism in the rat hepatoma. Texas reports on biology and Medicine 10: 273-278, 1952.
38. Dienes. L. and Zamecnik. P.C.: Transformation of bacteria into L forms by amino acids. J. Bact. 64:770-771, 1952.
39. Zamecnik, P.C.: Incorporation of radioactivity from DL-leucine-l- C14 into proteins of rat liver homogenates. Fed. Proc. 12: 295, 1953 (Abstract).
40. Keller, E.B. and Zamecnik, P.C.: Anaerobic incorporation of C14-amino acids into protein in cell-free liver preparations. Fed. Proc. 13: 239-240, 1954 (Abstract).
41. Zamecnik, P.C. and Keller, E.B.: Relation between phosphate energy donors and incorporation of labeled amino acids into proteins. J.Biol.Chem. 209: 337-354, 1954.
42. Keller, E.B., Zamecnik, P.C. and Loftfield, R.B.: The role of microsomes in the incorporation of amino acids into proteins. J. Histochem. and Cytochem. 2: 378-386, 1954.
43. Keller, E.B. and Zamecnik, P.C.: Effect of guanosine diphosphate on incorporation of labeled amino acids into proteins. Fed. Proc. 14: 234, 1955 (Abstract).
44. Littlefield, J.W., Keller, E.B., Gross, J, and Zamecnik, P.C.: Studies on cytoplasmic ribonucleoprotein particles from the liver of the rat. J.Biol.Chem. 217: 111-123, 1955.
45. Hoagland, M.B., Keller, E.B. and Zamecnik, P.C.: Enzymatic carboxyl activation of amino acids. J.Biol. Chem. 218: 345-358, 1956.
46. Zamecnik, P.C., Keller, E.B., Littlefield, .J.W., Hoagland. M.B. and Loftfield, R.B.: Mechanism of incorporation of labeled amino acids into proteins. J. Cell. And Comp. Physiol. 47, Supplement 1: 81-102, 1955.
47. Keller, E.B. and Zamecnik, P.C.: The effect of guanosine diphosphate and triphosphate on the incorporation of labeled amino acids into proteins. J Biol. Chem. 221: 45-59, 1956.
48. Stephenson, M,L. and Zamecnik, P.C.: Incorporation of C14-amino acids into proteins of leaf disks and cell-free fractions of tobacco leaves. Fed. Proc. 15: 1, 1956.
49. Stephenson, M.L., Thimann, K.V. and Zamecnik, P.C.: Incorporation of C14 amino acids into proteins of leaf disks and cell-free fractions of tobacco leaves. Arch Biochem. and Biophys. 65: 194-209, 1956.
50. Zamecnik, P.C., Keller, E.B,, Hoagland, M.B., Littlefield, J.W. and Loftfield, R,B.: Studies on the mechanism of protein synthesis. Ciba Foundation Symposium on Ionizing Radiations and Cell Metabolism. G.E.W. Wolstenholme and C.M. O'Connor, Editors, J. and A. Churchill, Ltd., London, 1956.
51 Hoagland, M.B., Zamecnik, P.C. and Stephenson, M.L.: Intermediate reactions in protein synthesis. Biochim. et Biophys. Acta 24: 215, 1957.
52. Zamecnik, P.C., Hoagland, M.B., and Stephenson, M.L.: Observations on the role of RNA in protein synthesis. Cellular Biology: Nucleic Acids and Viruses. Special Publications of the N.Y. Acad. Sci., Vol. V, pp 273.274, 1957.
53. Zamecnik, P.C., Stephenson, M.L., Scott, J.F. and Hoagland, M.B.: Incorporation of C14-ATP into soluble RNA isolated from 105,000 x g supernatant of rat liver. Fed. Proc. 16: 197, 1957.
54. Hoagland, M.B., Zamecnik, P.C. and Stephenson, M.L.: Intermediate reactions in protein biosynthesis. Fed. Proc. 16: 197, 1957.
55. Baker, W.H., Zamecnik, P.C. and Stephenson, M.L.: In vitro incorporation of C14-DL-1eucine into normal and leukemic white cells. Blood 12: 822-828, 1957.
56. Hoagland, M.B., Zamecnik, P.C., Sharon, N., Lipmann. F., Stulberg, M.P. and Boyer, P.D.: Oxygen transfer to AMP in the enzymatic synthesis of the hydroxamate of tryptophan. Biochim. et Biophvs. Acta 26: 215-217. 1957.
57. Zamecnik, P.C., Stephenson, M.L. and Hecht, L.I.: Intermediate reactions in amino acid incorporation. Proc. Natl. Acad. Sci. 44: 73-078, 1958.
58. Hoagland, M.B., Stephenson, M.L., Scott, J.F., Hecht, L.I, and Zamecnik, P.C.: A soluble ribonucleic acid intermediate in protein synthesis. J. Biol. Chem. 231: 241-256, 1958.
59. Hecht L.J., Stephenson, M.L. and Zamecnik, P.C.: Dependence of amino acid binding to soluble ribonucleic acid on cytidine triphosphate. Biochim. et Biophys. Acta 29: 460-461, 1958.
60. Zamecnik. P.C.: The microsomes. Sci. Am. 198: 118-124, 1958.
61. Hecht, L,J,, Zamecnik, P.C., Stephenson, M.L. and Scott, J.F.: Nucleoside triphosphates as precursors of ribonucleic acid end groups. J. Biol. Chem. 233: 954-963, 1958.
62. Stephenson, M.L., Hecht, L.I., Littlefield, J.W., Loftfield, R.B. and Zamecnik, P.C.: Intermediate reactions in protein synthesis. Symposium on Subcellular Particles, Am. Physiol. Soc., Washington, pp 160-171,1959.
63. Hoagland, M.B., Zamecnik, P.C. and Stephenson, M.L.: A hypothesis concerning the roles of particulate and soluble ribonucleic acids in protein synthesis. A Symposium on Molecular Biology, Zirkle, RE., Ed. Univ. of Chicago Press, pp 105-114. 1959.
64. Hecht, L.I., Stephenson, M.L. and Zamecnik, P.C.: Binding of amino acids to the end group of a soluble ribonucleic acid. Proc. Natl. Acad. Sci. 45: 505-518, 1959.
65. Zamecnik, P.C.: Historical and current aspects of the problem of protein synthesis. Harvey Lectures, Series 54 1958-1959, Academic Press, New York, pp 256-281, 1960.
66. Loftfield, R,B., Zamecnik, P.C., Hecht, L.E., Stephenson, M.L,, Hoagland, M.B., Littlefield, J.W. and Keller, E.B.: Studies on the mechanism of protein synthesis. 2nd UN Geneva Conference on Peaceful Uses of Atomic Energy. Proc. of Conf. 25: 121-124, Pergamon Press, London, 1959.
67. Zamecnik, P.C. and Stephenson, M.L.: Partial purification of transfer RNA. Ann. N.Y. Acad. Sci. 88:708-717,1960.
68. Lamborg, M.R., and Zamecnik, P.C.: Amino acid incorporation into protein by extracts of E.Coli. Biochim. et Biophys. Acta 42: 206-211, 1960.
69. Zamecnik, P.C., Stephenson, M.L. and Scott, J.F.: Partial purification of soluble RNA. Proc. Natl. Acad. Sci. 46: 811-822, 1960.
70. Zamecnik, P.C.: Soluble ribonucleic acid and protein synthesis. In The Molecular Control of Cellular Activity. McGraw-Hill Inc., NY, pp 259-264, 1961.
71. Zamecnik, P.C., Stephenson, M.L. and Yu, C.-T. Studies on preparation, fractionation and degradation of soluble ribonucleic acid. Symposium On Protein Biosynthesis, Wassenaar, The Netherlands, Academic Press, Inc., London, pp 125-131, Aug. 1960.
72. Yu, C.-T. and Zamecnik, P.C.: A hydrolytic procedure for ribonucleosides and its possible application to the sequential degradation of RNA. Biochem et Biophys. Acta 45:148-154, 1960.
73. Monier, R., Stephenson, M.L. and Zamecnik, P.C.: The preparation and some properties of a low molecu1ar weight ribonucleic acid from baker's yeast. Biochim et Biophys. Acta 41:1-8, 1960.
74. Zamecnik, P.C.: History and speculation on protein synthesis. Proc. of the Symposium on Mathematical Problems in the Biological Sciences, NY, Rand Corporation 14:47-53, 1962.
75. Stephenson, M,L. and Zamecnik, P.C.: Purification of valine transfer RNA by combined chromatographic and chemical procedures. Proc. Natl. Acad. Sci., 47:1627-1635, 1961.
76. Stephenson, M.L. and Zamecnik, P.C.: Isolation of valyl-RNA of a high degree of purity. Biodjern. Biophys. R~s. Comm. 7: 91-94, 1962.
77. Allen, D.W. and Zamecnik, P.C.: The effect of puromycin on rabbit reticulocyte ribbosomes. Biochim. et Biophys. Acta 55:865-874, 1962.
78. Zamecnik, P.C.: Unsettled questions in the field of protein synthesis. Jubilee lecture, Biochemical Society, London. Biochem. J. 85: 257-264, 1962.
79. Zamecnik, P.C.: DNA, RNA and Protein Synthesis. In Birth Defects, Fishbein, M., Ed. J.B. Ljppincott Co., Philadelphia, pp 106-113, 1963.
80. Zamecnik, P.C.: Aminoacyl ribonucleic acid and intermediary reactions in protein synthesis. Pontificia Academiae Scientiarum Scripta, Convegno internazionale sugli aspetti mederni della morfologia in biologia e in medicine. 1962, pp 432-437.
81. Zamecnik, P.C.: On the mechanism of protein synthesis. 5th Conf on Molecular Structure and Biochemical Reactions. Welch Foundation, Houston, Texas, Chapt. XII, 1962, pp 161-1 68.
82. Zamecnik, P.C.: Biosynthesis of protein and its relation to medical problems. Proc. of the 16th General Assembly of the Japan Medical Congress. Osaka, Japan, Vol. I, pp 269-274, 1963.
83. Allen, D.W. and Zamecnik, P.C.: T1 ribonuclease inhibition of polyuridylic acid-stimulated polyphenylalanine synthesis. Biochem. Biophys. Res. Comm. 11: 294-300, 1963.
84. Yu, C.-T. and Zamecnik, P.C.: On the amonoacyl-tRNA synthetase recognition sites of yeast and E. coli transfer RNA. Biochim Biophys. Res. Comm. 12: 457-463, 1963.
85. Yu, C.-T. and Zamecnik, P.C.: Effect on bromination on the amino acid accepting activities of transfer ribonucleic acid. Biochim. et Biophys.Acta 76: 209-222, 1963.
86. Yu, C.-T, and Zamecnik, P.C.: Effect of bromination on the biological activities of transfer RNA of E. coli. Science 144: 856-859, 1964.
87. Lamborg, M.R., Zamecnik, P.C., Li, T.-K., Kagi, J. and Vallee, B.L.: Anomalous rotatory dispersion of soluble ribonucleic acid and its relation to amino acid synthetase recognition. Biochemistry 4: 63-70, 1965.
88. Sarin, P.S. and Zamecnik, P.C.: On the stability of aminoacyl-s-RNA to nucleophilic catalysis. Biochim. et Biophys. Acta 91: 653-655, 1964.
89. Sarin, P.S. and Zamecnik, P.C.: Modification of amino acid acceptance and transfer capacity of s-RNA in the presence of organic solvents. Biochem. Biophys. Res. Comm. 19: 198-203, 1965.
90. Sarin, P.S. and Zamecnik, P.C.: Conformational differences between s-RNA and aminoacyl s-RNA. Biochem. Biophys. Res. Comm. 20: 400-405, 1965.
91. Lamborg, M.R. and Zamecnik, P.C.: Optical rotary dispersion of E. coli sRNA in the far ultraviolet region. Biochem. Biophys. Res. Comm. 20: 328-333, 1965.
92. Zamecnik, P.C.: The mechanism of protein synthesis and its possible alteration in the presence of oncogenic RNA viruses. (Presidential Address). Cancer Research 26: 1-6, 1966.
93. Sarin, P.S., Zamecnik, P.C., Bergquist, P.L. and Scott, J.F.: Conformational differences among purified samples of transfer RNA from yeast. Proc. Natl. Acad. Sci. 55: 579-585, 1966.
94. Zamecnik, P.C., Stephenson, M.L., Janeway, C.M. and Randerath, K.: Enzymatic synthesis of diadenosine tetraphosphate and diadenosine triphosphate with a purified lysyl-sRNA synthetase. Biochem. Biophys. Res. Com. 24: 91-97, 1966.
95. Randerath, K., Janeway, C.M., Stephenson, M.L. and Zamecnik, P.C.: Isolation and characterization of dinucleoside tetra- and triphosphates formed in the presence of lysyl-sRNA synthetase. Biochem. Biophys. Res. Comm. 24 98-105, 1966.
96. Zamecnik, P.C., Janeway, C.M., Randerath, K. and Stephenson, M.L.: A new family of dinucleotides whose formation is catalyzed by L-lysine:sRNA ligase (AMP). In Regulation of Nucleic Acid and Protein Biosynthesis, Koningsberger, V.V. and Bosch, L., Eds. Elsevier Pub. Co., Amsterdam, pp 169-176, 1967.
97. Zamecnik, P.C.: Presentation of the Kober Medal for 1966 to Joseph Charles Aub. Trans. Assoc. Am. Phys. 79: 84-98, 1966.
98. Zamecnik, P.C.: Concerning the recognition reaction and transfer RNA in protein synthesis. Symposium on Immunology and chemotherapy, Buffalo, Academic Press, NY, pp 155-166, 1967.
99. Zamecnik, M.V. and Zamecnik, P.C.: Mutation of chloroplasts in ageratum, following treatment with 5-bromodeoxyuridine. Exptl. Cell Research 45: 218-229, 1966.
100. Burton, K., Varney, N.F., and Zamecnik, P.C.: Action of osmium tetroxide on amino acid transfer ribonucleic acid. Correlations between the genetic code and the sensitivity of acceptor activity. Biochem. J. 99: 290-310, 1966.
101. Haines, J.A. and Zamecnik, P.C.: Chemical modification of aminoacyl ligases and the effect on formation of aminoacyl-tRNA. Biochim. et Biophys. Acta 146: 227-238, 1967.
102. Schofield, P. and Zamecnik, P.C.: Cupric ion catalysis in hydrolysis of aminoacyl-tRNA. Biochim. et Biophys. Acta 155: 410-416, 1968.
103. Zamecnik, P.C. and Stephenson, M.L.: A possible regulatory site located at the gateway to protein synthesis. In Regulatory Mechanisms for Protein Synthesis in Mammalian Cells, San Pietro, A., Lamborg, M.R. and Kenney, P.T., Eds. Academic Press, NY, pp 3-16, 1968.
104. Zamecnik, P.C. and Stephenson, M.L.: Nucleoside pyrophosphate related to the first step in protein synthesis. In Alfred Benzon Symposium I. The Role Of Nucleotides for the Function and Conformation of Enzymes, Kalckar, H.M., Klenow, H., Munch-Petersen, G., Ottesen, M. and Thuysen, J.M., Eds. Munksgaard, Copenhagen, pp 276-291, 1969.
105. Marshall, R.D. and Zamecnik, P.C.: Some physical properties of lysyl and arginyl transfer RNA synthetases of Escherichia Coli B. Biochim et Biopys. Acta 181: 454-464, 1969.
106. Zamecnik, P.C.: An historical account of protein synthesis with current overtones a personalized view. Cold Spring Harbor Symp. Quant. Biol. 34 1-16, 1969.
107. Scott, J.F. and Zamecnik, P.C.: Some optical properties of diadenosine-5'-phosphates. Proc. Natl. Acad. Sci. 64 1308-1314, 1969.
108. Marshall, R.D. and Zamecnik, P.C.: Aspects of the kinetic properties of lysyl-tRNA synthetase from E. Coli. strain B. Biochim. et Biophys. Acta 198: 376-385, 1970.
109. Bloemers, H.P.J., Stephenson, M.L, and Zamecnik, P.C.: A sensitive microassay for inorganic pyrophosphatase. Anal. Biochem. 34: 66-70, 1970.
110. Zamecnik, P.C. and Yu, C.-T.: Modification of tRNAs by bromination. In Methods in Enzymology, Col. 20, Moldave, K. and Grossman, L., Eds. Academic Press, NY, 1971, pp 175-178.
111. Zamecnik, P.C.: Summary of colloquium on "Transfer RNA and Transfer RNA Modification in Differentiation anbd Neoplasia." Cancer Research 31: 716-721, 1971.
112. Randerath, K., Rosenthal, L.J., and Zamecnik, P.C.: Base composition differences between avian myeloblastosis virus transfer RNA and transfer RNA isolated from normal and neoplastic host cells. Proc. Natl. Acad. Sci. 68: 3233-3237, 1971.
113. Zamecnik, P.C., Rosenthal, L.J. and Randerath, K.: Minor base changes in transfer ribonucleic acid in avian myeloblastosis virus. Proc. Vth Intl. Symp. on Comparative Leukemia Research, Padua, September 13-17. 1971.
114. Hirshfield, I.N. and Zamecnik, P.C.: Thiosine-resistant mutants of Escherichia coli K-l2 with growth-medium-dependent lysyl-tRNA synthetase activity. I. Isolation and physiological characterization. Biochim. et Biophys. Acta 259 330-343, 1972.
115. Hirshfield, I.N., Tanford, J.W. and Zamecnik, P.C.: Thiosine-resistant mutants of E. coli K-12 with growth-medium-dependent lysyl-tRNA synthetase activity. II. Evidence for an altered lysyl-tRNA synthetase. Biochim. et Biophys. Acta 259: 344-356, 1972.
116. Stephenson, M.L., Wirthlin, L.R.S., Scott, J.F. and Zamecnik, P.C.: An investigation of the 3'-terminal nucleocides of the high molecular weight RNA of avian myeloblastosis virus. Proc. Natl. Acad. Sci. USA 69: 1176-1180, 1972.
117. Long, J.C., Aisenberg, A.C., Zamecnik, M.V. and Zamecnik. P.C.: A tumor antigen in tissue cultures derived from patients with Hodgkin's disease. Proc. Natl. Acad. Sci. USA 70: 1540-1544, 1973.
118. Rosenthal, L.J, and Zamecnik, P.C.: Minor base composition of "70-S-associated" 4S RNA from avian myeloblastosis virus. Proc. Natl. Acad. Sci. USA 70: 865-869, 1973.
119. Rosenthal, L.J. and Zamecnik, P.C.: Amino acid acceptor activity of the "70-S-associated" 4S RNA from avian myeloblastosis virus. Proc. Natl. Acad. Sci. USA 70: 1184-1185, 1973.
120. Stephenson, M.L., Scott, J.F. and Zamecnik, P.C.: Evidence that the polyadenylic acid segment of "35S" RNA of avian myeloblastosis virus is located at the 3'-OH terminus. Biochem Biophys. Res. Comm. 55: 8-16, 1973.
121. Long, J.C., Aisenberg, A.C. and Zamecnik, P.C.: An antigen in Hodgkin's disease tissue cultures: radioiodine-labeled antibody studies. Proc. Natl. Acad. Sci. USA 71: 2605-2609, 1974. no abstract available.
122. Rapaport, E. and Zamecnik, P.C.: A new chemical procedure for 32-labeling of ribonucleic acids at their 5' ends after isolation. Proc. Natl. Acad. Sci. USA 72: 3124-317, 1975.
123. Rapaport, E., Svihovec, S. and Zamecnik, P.C.: Relationship of the first step in protein synthesis to ppGpp: Formation of A(5')ppp(5')Gpp. Proc. Natl. Acad. Sci. USA 72: 2653-2657, 1975.
124. Heine. U.I., Weber, G.H., Cottler-Fox, M., Layard, M.W., Stephenson, M.L. and Zamecnik, P.C.: Analysis of oncornavirus RNA subunits by electron microscopy. Proc. Natl. Acad. Sci. USA 72: 3716-3720, 1975.
125. Zamecnik, P.C., Joseph Charles Aub, 1890-1973. Trans. Assoc. Am. Physicians. 37 12-14, 1974.
126. Zamecnik. P.C.: Importance of South American non-human primates in human health and biomedical research. Keynote Address. Vii-x. Pan-American Health Organization, Lima, Peru, 2-4 June, 1975 (1976).
127. Zamecnik, P.C.: Protein synthesis early waves and recent ripples. In Reflections in Biochemistry, Kornberg, A., Horecker, B.L., Cornudella, L. and Oro, .J., Eds. Pergamon Prcss, N.Y., 1976, pp 303.308.
128. Hirshfield, I.N., Yeh, F.-M. and Zamecnik, P.C.: An in vivo effect of the metabolites L-alanine and glycyl-L-leucine on the properties of lysyl-tRNA synthetase from E. Coli K-12. I. Influence on subunit composition and molecular weight distribution. Biochim. et Biophys. Acta 435: 290-305, 1976.
129. Rapaport, E. and Zamecnik, P.C.: Incorporation of adenosine into ATP; Formation of compartmentalized ATP. Proc. Natl. Acad. Sci. USA 73: 3122.3125, 1976.
130. Rapaport, E. and Zamecnik, P.C.: The presence of diadenosine 5',5-pl, P4-tetraphosphate (Ap4A) in mammalian cells in levels varying widely with the proliferative activity of the tissue: A possible positive "pleiotypic activator". Proc. Natl. Acad. Sci. USA 73: 3984-3988, 1976.
131. Kolodny, M.H., Neville, A.C., Coleman, D.L. and Zamecnik, P.C.: Proton magnetic resonance investigation of interactions between L-tryptophan and dinucleoside phosphates at low pD. Biopolymers 16: 259, 1977.
132. Long, J.C., Aisenberg, A.C. and Zamecnik, P.C.: Chromatographic and electrophoretic analysis of an antigen in Hodgkin's disease tissue cultures. J. Natl. Cancer Inst. 58: 223-227, 1977.
133. Zamecnik, P.C. and Long, J.C.: Growth of cultured cells from patients with Hodgkin's disease and transplantation into nude mice. Proc. Natl. Acad. Sci. USA 74: 754-758, 1977.
134. Schwartz, D.E., Zamecnik, P.C. and Weith, H.L.: The Rous sarcoma virus genome is terminally redundant: The 3' sequence. Proc. Natl. Acad. Sci. USA 74: 994-998, 1977.
135. Long, J.C., Zamecnik, P.C., Aisenberg, A.C. and Atkins, L.: Tissue culture studies in Hodgkin's disease. Morphologic, cytogenetic, cell surface, and enzymatic properties of cultures derived from splenic tumors. J Exptl. Med. 145: 1484-1500, 1977. no abstract available.
136. Zamecnik, P.C. and Stephenson, M.L.: Inhibition of Rous sarcoma virus replication and transformation by a specific oligodeoxynucleotide. Proc. Natl. Acad. Sci. USA 75: 280-284, 1978.
137. Stephenson, M.L. and Zamecnik, P.C.: Inhibition of Rous sarcoma viral RNA translation by a specific oligodeoxyribonucleotide. Proc. Natl. Acad. Sci. USA 75: 285-288, 1978.
138. Rapaport, E. and Zamecnik, P.C.: Increased incorporation of adenosine into adenine nucleotide pools in serum-deprived mammalian cells. Proc. Natl. Acad. Sci. USA 75 1145-1147, 1978.
139. Zamecnik, P.C.: Historical aspects of protein synthesis. New York Academy of Science 325: 169-301, 1979.
140. Rapaport, E., Garcia-Blanco, M.A. and Zamecnik, P.C.: Regulation of DNA replication in S phase nuclei by ATP and ADP pools. Proc. Natl. Acad. Sci. USA 76: 1643, 1979. no abstract available. To order:
141. Plesner, P., Stephenson, M.L., Zamecnik, P.C. and Bucher, N.L.R.: Alfred Benzon Symposium, Munksgaard, Copenhagen. Diadenosine tetraphosphate (AP4A), an activator of gene function, Engberg, J. et al., Eds. Academic Press, NY, p 383, 1979.
142. Zamecnik, P.C.: Oligonucleotides and Growth Regulation. In Regulation of Macromoleular Synthesis of Low Molecular Weight Regulators, Koch, G. and Richter, D., Eds. Academic Press, NY, p 1-3, 1979.
143. Zamecnik, P.C. and Rapaport, E.: Historical Background on Diadenosine 5'5'''-p1p4 -tetraphosphate (Ap4A) and current developments. In Regulation of Macromolecular Synthesis by Low Molecular Weight Regulators, Koch, G. and Richter, D., Eds. Academic Press, NY, p 179-184, 1979.
144. Desselberger, U., Zamecnik, P. and Palese, P.: 3'-terminal sequences of hemagglutinin and neuraminidase genes of different Influenza A viruses. In Structure and Variation in Influenza Virus, Laver, G. and Air, G., Eds. Elsevier, NY, p 169-179, 1980.
145. Siekevitz, P. and Zamecnik, P.: Ribosomes and protein synthesis. In Discovery in Cell Biology, Siekevita, P., Porter, K. and Gall, J., Eds. Rockefeller U. Press, NY, 93, No. 3, Pt. 2, p 538-658, 1981.
146. Harris, N.L., Gang, D.I., Quay, S.C., Poppema, S., Zamecnik, P.C., Nelson-Rees, W.A. and O'Brien, S.J.: Contamination of Hodgkin's disease cell cultures. Nature 289:228-230, 1981.
147. Rapaport, E., Zamecnik, P.C. and Baril, E.F.: HeLa cell DNA polymerase a is tightly associated with tryptophanyl-tRNA synthetase and adenosine 5'5-p1,P4-tetraphosphate binding activities. Proc. Natl. Acad. Sci. USA 78: 838-842, 1981.
148. Rapaport, E., Zamecnik, P.C. and Baril, E.F.: Association of diadenosine 5'5-p',P4.tetraphosphate binding protein with HeLa cell polymerase-alpha. J.Biol Chem. 256: 12148-12151, 1981.
149. Zamecnik, P.C., Rapaport, E. and Baril, E.F.: Priming of DNA synthesis by diadenosine 5'5-p1,P4. tetraphosphate with a double-stranded octadecamer as a template and DNA polymerase-alpha. Proc. Natl. Acad. Sci. USA 79: 1791-1794, 1982.
150. Baril, E., Bonin, P., Burstein, D., Mara, K. and Zamecnik, P.: Resolution of the Ap4A binding subunit for a multiprotein form of HeLa cell DNA polynierase-alpha. Proc. Natl. Acad. Sci. USA 80: 4931-4935, 1983.
151. Holler, E., Holmquist, B., Vallee, B.L., Taneja, K. and Zamecnik, P.: Circular dichroism and secondary structure of bisnucleaoside oligophosphates and their Zn++ and Mg++ complexes. Biochemistry 22: 4924-4933, 1983.
152. Zamecnik, P.: Diadenosine 5'5'p1,P4-tetraphosphate (Ap4A): Its role in cellular metabolism. Analytical Biochem. 134: 1-10, 1983.
153. Elmaleh, D.R., Zamecnik, P.C., Castronovo, F.P., Jr., Strauss, H.W. and Rapaport. E.: 99"Tc-labeled nucleotides as tumor-seeking radiodiagnostic agents. Proc. Natl. Acad. Sci. USA 81: 918-921, 1984.
154. Zamecnik, P.: Historical and current studies on Ap4A. An orphan compound finds a home. Hoppe Seyler's Z. fur Physiol. Chem. Bd. 365: 610-611, 1984 (Abstract).
155. Flodgaard, H., Zamecnik, P.C., Meyers, K. and Klenow, H.: Ap4A determinations as a possible tool for the diagnosis of Chediak-Higashi disease and other platelet anomalies. Hoppe Seyler's Z. fr Physiol. Chem. Bd. 365: 610-6l1, 1984 (Abstract).
156. Chao, F.C. and Zamecnik, P.: Inhibition of platelet aggregation by Ap4A. Z. fur Physiol Chem. Bd. 365: 610, 1984 (Abstract).
157. Baril, E.F., Owen M.S., Vishwanatha, .J.K. and Zamecnik, P.C.: Ap4A interactions with a multiprotein form of DNA polymerase alpha primase from HeLa cells. Hoppe Seyler's Z. fur Physiol. Chem. Bd. 365: 614, 1984 (Abstract).
158. Zamecnik, P.: The machinery of protein synthesis. Biochemistry and the birth of molecular biology. TIBS. Letters 9: 464-466, 1984.
159. Rapaport, E., Yogeeswaran, G., Zamecnik, P.C. and Remy, P.: Covalent modification of phenylalanyl-tRNA synthetase with phenylalanine during the amino acid activation reaction catalyzed by the enzyme. J. Biol. Chem. 260: 9509-9512, 1985.
160. Kim, B.K., Chao, F.C., Leavitt, R., Fauci, A.S., Meyers, K.M. and Zamecnik, P.C.: Disadenosine 5',5 p1,P4-tetraphosphate deficiency in blood platelets of the Chediak-Higashi syndrome. B1ood. 66: 735-737, 1985.
161. Baril, E.F., Coughlin, S.A. and Zamecnik, P.C.: 5'5 p1,P4-diadenosine tetraphosphate (Ap4A): A putative initiator of DNA replication. Cancer Investigation 3: 465-471, 1985.
162. Flodgaard, H., Zamecnik, P.D., Meyers, K, and Klenow, H.: Diadenosine 5'5-p1,P4-tetraphosphate determinations in the diagnosis of Chediak-Higashi disease and other platelet dense granule anomalies. Thrombosis Research 37: 345-351, 1986.
163. Zamecnik, P.C., Goodchild, J., Taguchi, Y. and Sarin, P.S.: Inhibition of replication and expression of human T-cell lymphotropic virus type III in cultured cells by exogenous synthetic oligonucleotides complementary to viral RNA. Proc. Natl. Acad. Sci. USA 83: 4143-4146, 1987.
164. Plesner, P., Goodchild, J., Kalckar, H.M. and Zamecnik. P.C.: Oligonucleolides with rapid turnover of the phosphate groups occur endogenously in eukaryotic cells. Proc. Natl. Acad. Sci. USA 84: 1935-1939, 1987.
165. Rapaport, E., Remy, P., Kleinkauf, H., Vater, J., and Zamecnik, P.C.: Aminoacyl-tRNA synthetases catalyzed AMPADPATP exchange reactions, indicating labile covalent enzyme-amino acid intermediates. Proc. Natl. Acad. Sci. USA 84: 7891-7895, 1987.
166. Goodchild, J., Letsinger, R.L., Sarin, P.S., Zamecnik, M., and Zamecnik, P.C.: Inhibition of replication and expression of HIV-I in tissue culture by oligodeoxynucleotide hybridization competition in human retroviruses, cancer, and AIDS: Approach to prevention and therapy. UCLA Symposia on Molecular and Cellular Biology. New Series. Vol. 71. Bolognesi. 432-438. D, Ed. Alan Liss, Inc., NY. 1987
167. Louie, S., Kim, B.K. and Zamecnik, P.: Diadenosine 5' 5-p1,P4.Tetraphoshate, a potential antithrombotic agent. Thromb. Res. 49: 557-565, 1988.
168. Goodchild, J., Agrawal, S., Civeira, M., Sun, D., Sarin, P.S., and Zamecnik, P.C.: Inhibition of human immunodeficiency virus replication by antisense oligodeoxynucleotides. Proc. Natl. Acad. Sci. USA 85: 5507-5511,1988.
169. Agrawal, S., Goodchild, J., Civeira, M., Sarin, P., and Zamecnik, P.: The use of oligodeoxynucleotides for the inhibition of growth and expression of HIV in tissue culture in regulation of gene expression by RNA structure and anti-messengers. EMBO/Inserm Workshop, Les Arcs, Savoie, France, 2/28-3/5/88 (Abstract).
170. Agrawal, S., Goodchild, .J., Civeira, M.P., Thornton, A.H., Sarin, P.S. and Zamecnik, P.C.: Oligodeoxynucleoside phosphoramidates and phosphorothioates as inhibitors of human immunodeficiency virus. Proc. Natl. Acad. Sci. USA 85: 7079-7084, 1988.
171. Sarin, P.S., Agrawal, S., Civeira, M.P., Goodchild, J,, Ikeuchi, T., and Zamecnik, P.C.: Inhibition of acquired immunodeficiency syndrome virus by oligodeoxynucleoside methylphosphonates. Proc. Natl. Acad. Sci. USA 85: 7448.7451, 1988.
172. Cardullo, R.A., Agrawal, S., Flores, C., Zamecnik, P., and Wolf, D.E.: Detection of oligonucleotide hybridization by non-radiative and fluorescence resonance energy transfer. Proc. Natl. Acad. Sci. USA 85: 8790-8794,1988.
173. Goodchild, .J., Agrawal, S., Civeira, M.P., Sarin, P.S., Letsinger, R.L. and Zamecnik. P.C.: Antisense agents complementary to human immunodeficiency virus. Current Communications in Molecular Biology Antisense RNA and DNA. Cold Spring Harbor, NY, pp 135-139, 1988.
174. Agrawal, S., Ikeuchi, T., Sun, D., Sarin, P.S., Konopka, Z., Maizel, J. and Zamecnik, P.C.: Inhibition of Human Immunodeficiency Virus in Early-Infected and Chronically-Infected Cells by Antisense Oligodeoxynucleotides and their Phosphorothioate Analogues. Proc. Natl. Acad. Sci. USA 86 7790-7794, 1989.
175. Leiter, J.M., Agrawal, S., Palese, F. and Zamecnik, P.C.: Inhibition of influenza virus replication by phosphorothioate oligodeoxynucleotides. Proc. Natl. Acad. Sci. USA 87: 3430-3434, 1990.
176. Agrawal, S., Zamecnik, P.C.: Site specific functionalization of oligonucleotides for attaching two different reporter groups. Nucleic Acids Res. 18:5419-5423,
177. Agrawal, S., Sarin, P.S., and Zamecnik, P.C.: Inhibition of HIV replication and expression by synthetic oligonucleotides. J. Cell Biochem. 15D, p 10, 1990.
178. Zamecnik, P.C., and Agrawal, S.: The hybridization inhibition or antisense approach to the chemotherapy of AIDS. In Annual Review of AIDS Research. Wayne C., Koff, Wong Staal, F. and Kennedy, R.C., Eds. Marcel Dekker, pp 301-313, 1991.
179. Zamecnik, P.C.: Oligonucleotide base hybridization as a modulator of genetic message readout. In Advances in Antisense Nucleic Acid Therapy of Cancer and AIDS. pp 1-6, 1991.
180. Goodchild, J., Kim, B., and Zamecnik, P.: The clearance and degradation of oligodeoxynucleotides following intravenous injection into rabbits. Antisense Research and Development 1: 153-160, 1991.
182. Agrawal, S., Mayrand, S.H., Zamecnik, P.C. and Pederson, T.: Site-specific excision from RNA by Rnase H and mixed-phosphate-backbone oljgodeoxynucleotides. Proc. Natl. Acad. Sci. USA 87 1401-1405, 1990.
183. Agrawal, S., Sarin, P.S., Zamecnik, M. and Zamecnik, P.C.: Cellular uptake and anti-HIV activity of oligonucleotides and their analogues. In Gene Regulation by Antisense RNA and DNA, Erickson and Izant, Eds. Raven Press, NY, pp 273-283, 1992.
184. Zamecnik, P.C. and Agrawal, S.: Multiple roles of oligonucleotides in the regulation of gene expression. In New Leads and Targets in Drug Research. Krogsgaard-Larsen, P. et al. Eds. Munksgaard, Copenhagen, pp 48-59, 1991.
185. McLennan, A.G. and Zamecnik, P.C.: Dinucleoside polyphosphates Introduction. In Dinticleoside Phosphates in Metabolism. McLennan, AD.. Ed. CRC Press, Boca Raton, FL, pp 1-7, 1992.
186. Zamecnik, P.C., Kim, B., Guo, M.J., Taylor, G. and Blackburn, G.M.: Analogues of diadenosine 5'5 p1,p4-Tetraphosphate (AP4A) as potential anti-platelet-aggregation agents. Proc. Natl. Acad. Sci. USA 89: 2370-2373, 1992.
187. Rapaport E., Misiura, K., Agrawal, S. and Zamecnik, P.C.: Antimalarial activities of oligodeoxynucleotide phosphorothioates in chloroquine-resistant Plasmodium falciparum. Proc. Natl. Acad. Sci. USA 89: 8577-8580, 1992.
188. Sarin, P.S., Goldstein, A., Agrawal, S. and Zamecnik, P.: Use of drugs targeted to inhibit different stages of the HIV life cycle in the treatment of AIDS in combination therapies. In Biological Response Modifiers in the Treatment of Cancer and Infectious Diseases. Goldstein, A.L. and Garaci, E., Eds. Plenum Press, NY, pp 123-129, 1992.
189. Kim, B.K., Zamecnik, P., Taylor, G., Guo, M.J. and Blackburn, G.M.: Antithrombotic effects beta-beta'- monochloromethylene diadenosine 5'5_-P1,P4-tetraphosphate. Proc. Natl. Acad. Sci. USA 89: 11056-11058, 1992.
190. Lisziewicz, J., Sun, D., Klotman, M,, Agrawal, S., Zamecnik, P.C. and Gallo, R.: Specific inhibition of Human Immunodeficiency Virus Type I replication by antisense oligonucleotides; An in vitro model for treatment. Proc. Natl. Acad. Sci. USA 89: 11209-11213, 1992.
191. Agrawal, S., Tang, J., Sun. D., Sarin, P. and Zamecnik, P.C.: Synthesis and anti-HIV activity of oligoribonueleotides and their phosphorothioate analogues. Ann. NY Acad. Sci. 660: 2-10, 1992.
192. Lisziewicz, J., Sun, D., Metelev, V., Zamecnik, P., Gallo, R., and Agrawal, S.: Long-term treatment of human immunodeficiency virus-infected cells with antisense oligonucleotide phosphorothioates. Proc. Natl. Acad. Sci. USA 90: 3860-3864, 1993.
193. Nyilas, A., Agrawal, S. and Zamecnik, P.C.: Solid phase synthesis of oligonucleotides carrying puromycin at 3'-terminal. Bioorganic and Medicinal Chemistry Letters 3: 1371-1374, 1993.
194. Zamecnik, P., Aghajanian, J., Zamecnik, M., Goodchild, J., and Witman, G.: (1994). Electon micrographic studies of transport of oligodeoxynucleotides across eukaryotic cell membranes. Proc. Natl. Acad. Sci. USA, 91 3156-3160, 1994.
195. Temsamani, J., Metelev, V., Levina, A., Agrawal, S., and Zamecnik, P.: Inhibition of in vitro transcription by oligodeoxynucleotides. Antisense Res. And Devel. 4:279-284, 1994.
196. Zamecnik, P.: History of Antisense Oligonucleotides, in Antisense Oligonucleotides: Current Status: Ed. By Sudhir Agrawal, Humana Press, NJ, pp 1/11, 1996.
197. Zamecnik. P., Rapaport, E., Metelev, V., and Barker, R.: Inhibition of Drug-resistant P. falciparum in vitro by specific antisense phosphorothioate oligodeoxynucleotides. In Antisense Nucleic Acids. Ed. By R. Schlingensiepen, Blackwell Wissenschafts-Verlag, Berlin, Germany, 1997, pp 262-271.
198. Rapaport, E., Levina, A., Metelev, V., and Zamecnik, P.C.: Antimycobacterial Activities Of.Antisense Oligodeoxynucleotide Phosphorothioates in Drug-Resistant Strains. Proc. Natl. Acad. Sci. USA. 93:709-713, 1996.
199. Levina, A.S., Metelev, V.G., Cohen, A.S. and Zamecnik, P.C.: Conjugates of Minor Groove DNA binders with Oligodeoxynucleotides; Synthesis and Properties. Antisense Research and Development 6:75-85, 1996.
200. Barker, R.H., Jr., Metelev, V., Rapaport, E, and Zamecnik, P.: Inhibition of Plasmodium falciparum malaria using antisense oligodeoxynucleotides. Proc. Natl. Acad. Sci. USA 93:514-518, 1996.
201. Zamecnik. P.C.: Background of the antisense approach to chemotherapy. Antisense and Nucleic Acid Drug Development 7:199-202, 1997.
202. Chan, S.W., Gallo, S.J., Kim, B.K., Guo, M.J., Blackburn, G.M., and Zamecnik, P.C.: P1,P4.dithio-P2, P3-monochloromethylene diadenosine 5',5'''-P1,P4-tetraphosphate: a novel antiplatelet agent. Proc. Natl. Acad. Sci. USA 94:4034-4039, 1997.
203. Elmaleh, D.R., Narula, J., Babich, J.W., Petrov, A., Rapaport, E., Fischman, A.J., Khaw, Ban-An and Zamecnik, P.C.: Very early non-invasive detection of experimental atherosclerotic lesions with 99TClabeled diadenosince polyphosphates. Proc. Natl. Acad. Sci. USA 95:691-695, 1998.
204. Barker, R.H., Jr., Metelev. V., Coakley, A. and Zamecnik, P.C.: Effect of chemical structure on efficacy and specificity of antisense oligonucleotides against malaria in vitro. Exper. Parasitol. 88:51-59, 1998.
205. Harth, G., Zamecnik, P.C., Tang, J-Y, Tabatadze, D. and Horwitz, M.A.: Treatment of Mycobacterium tuberculosis with Antisense Oligonucleotides to Glutamine Synthetase, Formation of the poly-L-Glutamate/Glutamine Cell Wall Structure, and Bacterial Replication. Proc. Natl. Acad. Sci. USA 97:418-423, 2000
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Antisense therapeutics: Is it as simple as complementary base recognition? Sudhir Agrawal and Ekambar R. Kandimalla, Molecular Medicine Today, 2000, 6:2:72-81.
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Antisense therapeutics, Sudhir Agrawal, Qiuyan Zhao, Current Opinion in Chemical Biology 1998, 2:519-528.
Sensible use of antisense: how to use oligonucleotides as research tools, Kathleen J. Myers and Nicholas M. Dean, Trends in Pharmacological Sciences, 2000, 21:1:19-23.
Antisense oligonucleotides: strategies for delivery, Keith J. Miller and Sudip K. Das, Pharmaceutical Science and Technology Today, 1998, 1:9:377-386.
Antisense oligonucleotides: a systematic high-throughput approach to target validation and gene function determination, Margaret F. Taylor, Kristin Wiederholt and Fran Sverdrup, Drug Discovery Today, 1999, 4:12:562-567.
Why antisense technology makes good sense for cancer treatment, Kathryn Senior, Pharmaceutical Science and Technology Today, 2000, 3:7:217-218.
Coding in the Noncoding DNA Strand: A Novel Mechanism of Gene Evolution, J Mol Evol 2000 Dec;51(6):600-606, Boldogkoi Z. Laboratory of Neuromorphology, Department of Anatomy, Semmelweis University Budapest, Hungary. The question whether the noncoding DNA strand had or still has the capability for encoding functional polypeptides has been addressed in several articles. The theoretical background of the views advocating this idea arose from two groups of findings. One of them was based on various observations implying that the genetic code was adapted for double-strand coding. The other group of theories arose from the observation of gene-length overlapping open reading frames (O-ORFs) on the antisense DNA strand in a number of genes. In fact, the above theories, which I term selectionist, conceive a novel conception of gene evolution, proposing that new genes can be created by the utilization of antisense DNA strand. In contrast, neutralist theory claims that the O-ORFs are mere by-products of evolutionary processes acting to create special codon usage and base distribution patterns in the coding sequences.
Suppression of gene expression by targeted disruption of messenger RNA: available options and current strategies., Stem Cells 2000;18(5): 307-19, Jen KY, Gewirtz AM, Department of Cell and Molecular Biology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania. At least three different approaches may be used for gene targeting including: A) gene knockout by homologous recombination; B) employment of synthetic oligonucleotides capable of hybridizing with DNA or RNA, and C) use of polyamides and other natural DNA-bonding molecules called lexitropsins. Targeting mRNA is attractive because mRNA is more accessible than the corresponding gene. Three basic strategies have emerged for this purpose, the most familiar being to introduce antisense nucleic acids into a cell in the hopes that they will form Watson-Crick base pairs with the targeted gene's mRNA. Duplexed mRNA cannot be translated, and almost certainly initiates processes which lead to its destruction. The antisense nucleic acid can take the form of RNA expressed from a vector which has been transfected into the cell, or take the form of a DNA or RNA oligonucleotide which can be introduced into cells through a variety of means. DNA and RNA oligonucleotides can be modified for stability as well as engineered to contain inherent cleaving activity. It has also been proven that because RNA and DNA are very similar chemical compounds, DNA molecules with enzymatic activity could also be developed. This assumption proved correct and led to the development of a "general-purpose" RNA-cleaving DNA enzyme. The attraction of DNAzymes over ribozymes is that they are very inexpensive to make and that because they are composed of DNA and not RNA, they are inherently more stable than ribozymes. Although mRNA targeting is impeccable in theory, many additional considerations must be taken into account in applying these strategies in living cells, including mRNA site selection, drug delivery and intracellular localization of the antisense agent. Nevertheless, the ongoing revolution in cell and molecular biology, combined with advances in the emerging disciplines of genomics and informatics, has made the concept of nontoxic, cancer-specific therapies more viable then ever and continues to drive interest in this field.
Antisense and nuclear medicine., J Nucl Med 1999 Apr;40(4):693-703, Hnatowich DJ, Department of Nuclear Medicine, University of Massachusetts Medical Center, Worcester. Despite many uncertainties concerning mechanism, synthetic single-strand antisense deoxyribonucleic acids (DNAs) are now in clinical trials for the chemotherapy of viral infections such as human immunodeficiency virus (HIV) and human papilloma virus; several cancers, including follicular lymphoma and acute myelogenous leukemia; inflammatory processes such as Crohn's disease and rheumatoid arthritis; and in allergic disorders. The expectation is that antisense DNAs will be important to future chemotherapy. The question considered here is whether antisense DNAs will also be important to future nuclear medicine imaging. While efforts toward developing antisense imaging are comparatively nonexistent thus far, investigations into the mechanisms of cellular transport and localization and the development of a second generation of antisense DNAs have occurred largely within the antisense chemotherapy industry. Fortunately, many of the properties of DNA for antisense imaging, such as high in vivo stability and adequate cell membrane transport, are the same as those for antisense chemotherapy. Unfortunately, interests diverge in the case of several other key properties. For example, rapid localization and clearance kinetics of the radiolabel and prolonged retention in the target are requirements unique to nuclear medicine. No doubt the development of antisense imaging will continue to benefit from improvements in the antisense chemotherapy industry. However, a considerable effort will be required to optimize this approach for imaging (and radiotherapy). The potential of specifically targeting virtually any disease or normal tissue should make this effort worthwhile.
Potent and nontoxic antisense oligonucleotides containing locked nucleic acids, Wahlestedt C, Salmi P, Good L, Kela J, Johnsson T, Hokfelt T, Broberger C, Porreca F, Lai J, Ren K, Ossipov M, Koshkin A, Jakobsen N, Skouv J, Oerum H, Jacobsen MH, Wengel J, Proc Natl Acad Sci U S A 2000 May 9;97(10):5633-8, Center for Genomics Research and Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden. Insufficient efficacy and/or specificity of antisense oligonucleotides limit their in vivo usefulness. We demonstrate here that a high-affinity DNA analog, locked nucleic acid (LNA), confers several desired properties to antisense agents. Unlike DNA, LNA/DNA copolymers were not degraded readily in blood serum and cell extracts. However, like DNA, the LNA/DNA copolymers were capable of activating RNase H, an important antisense mechanism of action. In contrast to phosphorothioate-containing oligonucleotides, isosequential LNA analogs did not cause detectable toxic reactions in rat brain. LNA/DNA copolymers exhibited potent antisense activity on assay systems as disparate as a G-protein-coupled receptor in living rat brain and an Escherichia coli reporter gene. LNA-containing oligonucleotides will likely be useful for many antisense applications.
Inhibition of human cancer cell growth by inducible expression of human ribonucleotide reductase antisense cDNA., Antisense Nucleic Acid Drug Dev 2000 Apr;10(2):111-6, Chen S, Zhou B, He F, Yen Y, Cancer Center, Veterans General Hospital, National Yang-Ming University Taipei, Taiwan. Ribonucleotide reductase (RR) is a rate-limiting enzyme in DNA synthesis and repair. The enzyme consists of two dissimilar subunits, M1 and M2. It is known that the M2 subunit plays a role in tumorigenicity and metastasis. In this study, we transfected human oropharyngeal KB cancer cells with human RR M1 and M2 antisense cDNA expressed by an inducible vector system. The transfectants were double-selected with hygromycin and G418. The clones, designated KB-M1AS, KB-M2AS and KB-CAT, represented transfectant clones that contained M1 antisense cDNA, M2 antisense cDNA, and a CAT reporter gene, respectively. In a colony-forming assay, colony formation for the KB-M2AS clone decreased approximately 50 percent when M2 antisense mRNA expression was induced by isopropylthiogalactose (IPTG). However, the KB-M1AS clone revealed no significant inhibition under IPTG induction. RR enzyme activity, as measured by 14CDP reduction assay, revealed a 30 percent decrease in the IPTG-induced KB-M2AS clone relative to non-IPTG-induced samples at 144 hours. As shown by Northern blot, expression of the M2 antisense mRNA showed peaks at 48 hours and 144 hours after induction by IPTG. M2 antisense mRNA expression induced by IPTG was 33-fold greater than the uninduced control at 144 hours. Western blot analysis showed that the M2 subunit protein level decreased in the KB-M2AS clone beginning at 72 hours after induction and continued to decrease to 50 percent of the uninduced control at 144 hours, then showed a slight recovery at 168 hours. In conclusion, M2 antisense mRNA expression by an inducible system can effectively decrease RR M2 protein expression, reduce enzyme activity, and inhibit growth. Furthermore, this approach can be employed in future antisense investigations.
A Rodent Model of Protein Turnover Used to Design an Experiment for Measuring the Rates of Channeling, Recycling and Protein Synthesis., J Nutr 2000 Dec;130(12):3097-3102, Johnson HA, Baldwin RL, Klasing KC, France J, Calvert CC, Animal Science Department, University of California at Davis, and The University of Reading, Department of Agriculture, Earley Gate, Reading, UK. We described previously a mechanistic model of whole-body protein turnover in rodents. Channeling was defined as the flow of amino acids from the extracellular compartment to aminoacyl tRNA and protein synthesis. Recycling was defined as the flow of amino acids from protein degradation to aminoacyl tRNA (protein synthesis) without mixing with the intracellular pool of amino acids. In this paper, the model is applied to tissues and whole body and is used to develop an experimental protocol for estimating protein fractional synthesis rate, recycling and channeling. Channeling, recycling and protein synthesis must be estimated simultaneously because changes in specific radioactivities over time are highly dependent on the rate of protein synthesis. Injection-specific radioactivities, body weights and experimental variation were used with the model to generate data at different rates of recycling and channeling. The data generated were then used to determine the best time points and experimental method to estimate percentages of recycling, channeling and protein synthesis rate by the iterative Method of Maximum Likelihood. Specific radioactivity at each time point was based on simulated data from three rodents at each of six time points. Predicted protein synthesis rates were within 5 percent/d of observed rates for all methods. Predicted rates of recycling and channeling were generally within 15 percent of observed rates except recycling in muscle at high channeling and high recycling. Standard deviations of the predictions of percentages of channeling and recycling were between 0.148 and 44.5 percent for the pulse dose method, 0.0655 and 197 percent for the continuous infusion method and 0.351 and 962 percent for the flooding dose method. The experimental design that yields the best estimates of channeling, recycling and protein synthesis is the pulse dose. Changes in amino acid specific radioactivities in the extracellular, aminoacyl tRNA and protein pools were greatest and should be measured at 2, 6, 10, 40, 70 and 100 min in the pulse method.
Transcription of Eukaryotic Protein-Coding Genes, Annu Rev Genet 2000;34:77-137, Lee TI, Young RA, Department of Biology, Massachusetts Institute of Technology, Cambridge. The past decade has seen an explosive increase in information about regulation of eukaryotic gene transcription, especially for protein-coding genes. The most striking advances in our knowledge of transcriptional regulation involve the chromatin template, the large complexes recruited by transcriptional activators that regulate chromatin structure and the transcription apparatus, the holoenzyme forms of RNA polymerase II involved in initiation and elongation, and the mechanisms that link mRNA processing with its synthesis. We describe here the major advances in these areas, with particular emphasis on the modular complexes associated with RNA polymerase II that are targeted by activators and other regulators of mRNA biosynthesis.
Inhibition of Huntingtin synthesis by antisense oligodeoxynucleotides., Nellemann C, Abell K, Norremolle A, Lokkegaard T, Naver B, Ropke C, Rygaard J, Sorensen SA, Hasholt L, Mol Cell Neurosci 2000 Oct;16(4):313-23, Department of Medical Biochemistry and Genetics, Panum Institute, University of Copenhagen, Denmark. The Huntington disease gone encodes the protein huntingtin, which is widely expressed during embryonic development and in mature tissues. In order to elucidate the physiological function of huntingtin, which so far is unknown, we intend to study the effect of antisense down-regulated huntingtin expression. We have found an inhibiting effect of a phosphorothioated oligodeoxynucleotide (PS-ODN) added to the culture medium of embryonic teratocarcinoma cells (NT2) and postmitotic neurons (NT2N neurons) differentiated from the NT2 cells. Specific inhibition of expression of endogenous huntingtin was achieved in NT2N neurons in the concentration range of 1-5 microM PS-ODN, whereas no inhibition was obtained in NT2 cells. We describe in detail the selection of the target sequence for the antisense oligo and the uptake, intracellular distribution, and stability of the antisense PS-ODN in the two cell types. Antisense down-regulation of huntingtin in this model of human neurons represents a suitable approach to study its normal function.
Domain-specific recruitment of amide amino acids for protein synthesis., Tumbula DL, Becker HD, Chang WZ, Soll D, Nature 2000 Sep 7;407(6800):106-10, Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut. The formation of aminoacyl-transfer RNA is a crucial step in ensuring the accuracy of protein synthesis. Despite the central importance of this process in all living organisms, it remains unknown how archaea and some bacteria synthesize Asn-tRNA and Gln-tRNA. These amide aminoacyl-tRNAs can be formed by the direct acylation of tRNA, catalysed by asparaginyl-tRNA synthetase and glutaminyl-tRNA synthetase, respectively. A separate, indirect pathway involves the formation of mis-acylated Asp-tRNA(Asn) or Glu-tRNA(Gln), and the subsequent amidation of these amino acids while they are bound to tRNA, which is catalysed by amidotransferases. Here we show that all archaea possess an archaea-specific heterodimeric amidotransferase (encoded by gatD and gatE) for Gln-tRNA formation. However, Asn-tRNA synthesis in archaea is divergent: some archaea use asparaginyl-tRNA synthetase, whereas others use a heterotrimeric amidotransferase (encoded by the gatA, gatB and gatC genes). Because bacteria primarily use transamidation, and the eukaryal cytoplasm uses glutaminyl-tRNA synthetase, it appears that the three domains use different mechanisms for Gln-tRNA synthesis; as such, this is the only known step in protein synthesis where all three domains have diverged. Closer inspection of the two amidotransferases reveals that each of them recruited a metabolic enzyme to aid its function; this provides direct evidence for a relationship between amino-acid metabolism and protein biosynthesis.
The adaptor hypothesis revisited., Ibba M, Becker HD, Stathopoulos C, Tumbula DL, Soll D, Trends Biochem Sci 2000 Jul;25(7):311-6, Center for Biomolecular Recognition, Dept of Medical Biochemistry and Genetics, Laboratory B, The Panum Institute, Copenhagen, Denmark. As originally postulated in Crick's Adaptor hypothesis, the faithful synthesis of proteins from messenger RNA is dependent on the presence of perfectly acylated tRNAs. The hypothesis also suggested that each aminoacyl-tRNA would be made by a unique enzyme. Recent data have now forced a revision of this latter point, with an increasingly diverse array of enzymes and pathways being implicated in aminoacyl-tRNA synthesis. These unexpected findings have far-reaching implications for our understanding of protein synthesis and its origins.
An easy cell-free protein synthesis system dependent on the addition of crude Escherichia coli tRNA., Kanda T, Takai K, Yokoyama S, Takaku H, J Biochem (Tokyo) 2000 Jan;127(1):37-41, Department of Industrial Chemistry, Chiba Institute of Technology, Tsudanuma, Narashino, Chiba, Japan. The protein-synthesizing S30 extract of Escherichia coli contains tRNA, which limits its applications in cell-free protein synthesis. Here, we show that at least Arg- and Ser-acceptor activities can be removed from a standard S30 extract by treatment with an immobilized RNase A resin. This RNase-treated extract exhibits no protein synthesis activity, but regains it when supplied with crude E. coli tRNA and a small amount of human placental RNase inhibitor. The protein synthesis is dependent on the addition of tRNA in the presence of the RNase inhibitor. Chloramphenicol acetyltransferase was synthesized with this system and found to be active.
Quality control mechanisms during translation., Ibba M, Soll D, Science 1999 Dec 3;286(5446):1893-7, Center for Biomolecular Recognition, Department of Medical Biochemistry and Genetics, Laboratory B, Panum Institute, Copenhagen, Denmark. Translation uses the genetic information in messenger RNA (mRNA) to synthesize proteins. Transfer RNAs (tRNAs) are charged with an amino acid and brought to the ribosome, where they are paired with the corresponding trinucleotide codon in mRNA. The amino acid is attached to the nascent polypeptide and the ribosome moves on to the next codon. The cycle is then repeated to produce a full-length protein. Proofreading and editing processes are used throughout protein synthesis to ensure the faithful translation of genetic information. The maturation of tRNAs and mRNAs is monitored, as is the identity of amino acids attached to tRNAs. Accuracy is further enhanced during the selection of aminoacyl-tRNAs on the ribosome and their base pairing with mRNA. Recent studies have begun to reveal the molecular mechanisms underpinning quality control and go some way to explaining the phenomenal accuracy of translation first observed over three decades ago.
Transfer RNA-dependent translocation of misactivated amino acids to prevent errors in protein synthesis., Nomanbhoy TK, Hendrickson TL, Schimmel P, Mol Cell 1999 Oct;4(4):519-28, Skaggs Institute for Chemical Biology, Scripps Research Institute, Beckman Center, La Jolla, California. Misactivation of amino acids by aminoacyl-tRNA synthetases can lead to significant errors in protein synthesis that are prevented by editing reactions. As an example, discrete sites in isoleucyl-tRNA synthetase for amino acid activation and editing are about 25 A apart. The details of how misactivated valine is translocated from one site to the other are unknown. Here, we present a kinetic study in which a fluorescent probe is used to monitor translocation of misactivated valine from the active site to the editing site. Isoleucine-specific tRNA, and not other tRNAs, is essential for translocation of misactivated valine. Misactivation and translocation occur on the same enzyme molecule, with translocation being rate limiting for editing. These results illustrate a remarkable capacity for a specific tRNA to enhance amino acid fine structure recognition by triggering a unimolecular translocation event.
How translational accuracy influences reading frame maintenance., Farabaugh PJ, Bjork GR, EMBO J 1999 Mar 15;18(6):1427-34, Department of Biological Sciences, University of Maryland Baltimore County, Baltimore. Most missense errors have little effect on protein function, since they only exchange one amino acid for another. However, processivity errors, frameshifting or premature termination result in a synthesis of an incomplete peptide. There may be a connection between missense and processivity errors, since processivity errors now appear to result from a second error occurring after recruitment of an errant aminoacyl-tRNA, either spontaneous dissociation causing premature termination or translational frameshifting. This is clearest in programmed translational frameshifting, where the mRNA programs errant reading by a near-cognate tRNA; this error promotes a second frameshifting error (a dual-error model of frameshifting). The same mechanism can explain frameshifting by suppressor tRNAs, even those with expanded anticodon loops. The previous model that suppressor tRNAs induce quadruplet translocation now appears incorrect for most, and perhaps for all of them. We suggest that the "spontaneous" tRNA-induced frameshifting and "programmed" mRNA-induced frameshifting use the same mechanism, although the frequency of frameshifting is very different. This new model of frameshifting suggests that the tRNA is not acting as the yardstick to measure out the length of the translocation step. Rather, the translocation of 3 nucleotides may be an inherent feature of the ribosome.
Removing tRNA from a cell-free protein synthesis system for use in protein production., Kanda T, Takai K, Yokoyama S, Takaku H; Nucleic Acids Symp Ser 1997;(37):319-20; Department of Industrial Chemistry, Chiba Institute of Technology, Japan. The cell-free system for biosynthesis of proteins is becoming an important tool for protein engineering. In particular, introduction of the unnatural amino acids is achieved though cell-free protein synthesis with the use of chemically acylated tRNA that recognizes a specific codon. In the original method, however, it was difficult to control the system through changing tRNA composition, as the endogenous tRNAs are involved in the reaction. Thus, in the present study, we digested the tRNA within Escherichia coli S30 extract with resin-bound RNase A, and estimated the protein synthesis activity. It was revealed that this digestion process does not damage the activity, if a protease inhibitor, phenylmethylsulfonyl fluoride (PMSF), is present in the digestion reaction.
Regulation of protein synthesis by minigene expression., Hernandez J, Ontiveros C, Valadez JG, Buckingham RH, Guarneros G, Biochimie 1997 Sep;79(8):527-31, Departamento de Genetica y Biologia Molecular, Centro de Investigacion y de Estudios Avanzados del IPN, Mexico City, Mexico. Peptidyl-tRNA hydrolase (Pth), an enzyme essential for Escherichia coli viability, scavenges peptidyl-tRNA released during abortive polypeptide chain elongation. Bacterial strains of E. coli partially defective in Pth activity are unable to maintain bacteriophage lambda growth. Phage mutations that overcome the bacterial defect have been located to several regions in the lambda genome named bar. Plasmid constructs expressing just the bar region are toxic and cause a general arrest of protein synthesis in Pth-defective cells. Inspection of the nucleotide sequence from two bar regions reveals the short coding sequence AUG AUA Stop, spaced by an AT-rich segment from a Shine Dalgarno-like sequence (S-D). These sequences have been named minigenes. Base changes altering the putative S-D, the two sense codons, or the stop codon have been found to reduce Bar-toxicity. Transcripts containing bar function as mRNA. Upon expression in pth mutants, wild-type (bar+) transcripts are found associated with ribosomes. In addition, bar+ RNA forms ternary complexes with the 30S ribosomal subunit and the initiator tRNA and can be released upon run-off translation in the same way as an authentic mRNA. A cell free system for protein synthesis reproduces the in vivo effects: bar+ expression inhibits protein synthesis, bar+ RNA sequences are associated with ribosomes in the inhibited extracts, addition of purified Pth restores synthesis, and excess of tRNA(Lys), specific for the last sense codon in a mutant toxic minigene, prevents protein synthesis inhibition. Also, bar expression promotes association of methionine with ribosomes possibly in a translation complex. These results are consistent with a model proposing tRNA starvation to explain the behaviour of a pth mutant, thermosensitive for protein synthesis.
The origin of the genetic code and protein synthesis., Alberti S, J Mol Evol 1997 Oct;45(4):352-8, Department of Cell Biology and Oncology, Istituto di Ricerche Farmacologiche Mario Negri, Consorzio Mario Negri Sud, Chieti, Italy. A model for a parallel evolution of the genetic code and protein synthesis is presented. The main tenet of this model is that the genetic code—that is, a correspondence between nucleotide and amino-acid coding units—originated from sequence-specific interaction between abiotically synthesized polynucleotides and polypeptides. A sequence-specific binding between oligonucleotides and oligopeptides is supported by experimental findings. Moreover, it is parsimonious enough to be consistent with the relatively simple chemistry of a primordial environment. Proximity between peptides and RNA increased the rate of formation of ester bonds between them. This lead to the accumulation of sequence-specific polypeptide-polynucleotide pairs—that is, of primordial-loaded tRNA. Condensation of short polypeptides into longer products could be catalyzed by a sequence-specific juxtaposition of loaded tRNA over complementary RNA, originating the core of protein synthesis. The accumulation of useful encoded products—for example, catalysts for tRNA loading (primordial aminoacyl-tRNA synthetases) or stabilizers of tRNA-mRNA interactions (primordial ribosomes)—permitted the subsequent evolution of protein synthesis and of the genetic code to their mature form. This occurred via a parallel reduction in length of the interacting polynucleotides and polypeptides. Thus, it maintained the correct reading frame of mRNA from the preceding stages of evolution.
Synthesis of the translational apparatus is regulated at the translational level., Meyuhas O, Eur J Biochem 2000 Nov;267(21):6321-30, Department of Biochemistry, The Hebrew University-Hadassah Medical School, Jerusalem, Israel. The synthesis of many mammalian proteins associated with the translational apparatus is selectively regulated by mitogenic and nutritional stimuli at the translational level. The apparent advantages of the regulation of gene expression at the translational level are the speed and the readily reversible nature of the response to altering physiological conditions. These two features enable cells to rapidly repress the biosynthesis of the translational machinery upon shortage of amino acids or growth arrest, thus rapidly blocking unnecessary energy wastage. Likewise, when amino acids are replenished or mitogenic stimulation is applied, then cells can rapidly respond in resuming the costly biosynthesis of the translational apparatus. A structural hallmark, common to mRNAs encoding many components of the translational machinery, is the presence of a 5' terminal oligopyrimidine tract (5'TOP), referred to as TOP mRNAs. This structural motif comprises the core of the translational cis-regulatory element of these mRNAs. The present review focuses on the mechanism underlying the translational control of TOP mRNAs upon growth and nutritional stimuli. A special emphasis is put on the pivotal role played by ribosomal protein S6 kinase (S6K) in this mode of regulation and the upstream regulatory pathways, which might be engaged in transducing external signals into activation of S6K. Finally, the possible involvement of pyrimidine-binding proteins in the translational control of TOP mRNAs is discussed.
Regulation of mammalian gene expression., Beyersmann D, EXS 2000;89:11-28, Department of Biology and Chemistry, University of Bremen, Germany. The expression of mammalian genes is regulated primarily at the level of initiation of transcription. The regulatory structure of the mammalian genes consists of the sequence coding for a protein, a proximal upstream promoter sequence which binds the general (basal) transcription factors, and a distant enhancer sequence which binds the inducible transcription factors. The general transcription factors are proteins which combine with the RNA polymerase at the promoter to form the initiation complex. Binding of the inducible transcription factors at short DNA sequences, named response elements, mostly enhances or rarely represses the formation of the initiation complex. Transcription factors share common structural motifs; the most frequent are zinc finger, leucine zipper and helix-loop-helix structures. Inducible transcription factors are activated to bind their target response elements on DNA by protein kinases, by binding of activating or removal of inhibitory factors, or by de novo protein synthesis. Inducible transcription factors are activated by hormones or growth factors addressing a number of genes which share common response elements. Steroid and thyroid hormones combine with intracellular receptors to form active transcription factors. Other transcription factors are activated by protein kinases which are themselves activated by hormones through cell membrane receptors and further cellular signaling paths. Whereas the main level of transcriptional control is the initiation of RNA synthesis, in some instances genes are also regulated by alternative splicing of the primary transcript or control of translation into proteins. Large-scale silencing of genes is mediated by the packing of DNA in highly condensed heterochromatin structures and DNA methylation at cytosines in defined guanine-cytosine (GC)-sequences.