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Specific binding of tryptophan transfer RNA to avian myeloblastosis virus RNA-dependent DNA polymerase (reverse transcriptase)

Panet, Amos and Haseltine, William A. and Baltimore, David and Peters, Gordon and Harada, Fumio and Dahlberg, James E. (1975) Specific binding of tryptophan transfer RNA to avian myeloblastosis virus RNA-dependent DNA polymerase (reverse transcriptase). Proceedings of the National Academy of Sciences of the United States of America, 72 (7). pp. 2535-2539. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:PANpnas75

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Abstract

The ability of tryptophan tRNA (tRNATrp) to initiate reverse transcription of the 70S RNA of avian RNA tumor viruses suggested that the reverse transcriptase (RNA-dependent DNA polymerase; deoxynucleosidetriphosphate: DNA deoxynucleotidyltransferase; EC 2.7.7.7) might have a specific binding site for the tRNA. A complex of tRNATrp and the avian myeloblastosis virus reverse transcriptase has been demonstrated using chromatography on Sephadex G-100 columns. Of all the chicken tRNA's, only tRNATrp and a tRNA4{}Met bind to the enzyme with high enough affinity to be selected from a mixture of the chicken cell tRNAs. The ability of tRNATrp to change the sedimentation rate of the enzyme indicates that tRNATrp is not binding to a contaminant in the enzyme preparation. Treatment of the enzyme with monospecific antibody to reverse transcriptase prevented binding of tRNA as well as inhibited the DNA polymerase activity of the enzyme. The ability of reverse transcriptase to utilize tRNATrp as a primer for DNA synthesis, therefore, appears to involve a highly specific site on the enzyme.


Item Type:Article
Additional Information:© 1975 by the National Academy of Sciences. Contributed by David Baltimore, April 7, 1975. We are grateful to Drs. U. L. RajBhandary and H. M. Temin for helpful advice and the provision of materials and to B.P. Stark for excellent technical assistance. This work was supported by a contract from the Virus Cancer Program of the National Cancer Institute, Grant no. CA-14051 from the National Cancer Institute, and Grants no. CA-15166 from the National Institutes of Health and no. GB-32152X from the National Science Foundation. W.H. was a fellow of the Helen Hay Whitney Foundation. D.B. was a Research Professor of the American Cancer Society. J.E.D. was supported by a Research Career Development Award (no. GM-32770) from the National Institutes of Health.
Subject Keywords:primer; DNA synthesis; 70S RNA; RNA tumor virus
Record Number:CaltechAUTHORS:PANpnas75
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:PANpnas75
Alternative URL:http://dx.doi.org/10.1073/pnas.72.7.2535
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:7630
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:15 Mar 2007
Last Modified:26 Dec 2012 09:33

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