J Virol. 1992 July; 66(7): 4271-4278
When retroviral reverse transcriptases reach the end of their RNA templates.
T B Fu and
J Taylor
Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111.
ABSTRACT
Luo and Taylor (J. Virol. 64:4321-4328, 1990) have previously shown that when, during RNA-directed DNA synthesis, a retroviral reverse transcriptase comes to a halt at the end of an RNA template, the associated RNase H produces a specific oligonucleotide that contains the 5' end of that template; in those studies the length of the oligonucleotide was predominantly 17 nucleotides. We have now investigated variables that might affect the formation and length of such a terminal oligonucleotide. We found small but significant variations in the length could be caused by the choice of reaction conditions and also the sources of reverse transcriptase and RNA template. Nevertheless, the general finding in all these situations was that RNase H acted at or about 14 to 18 nucleotides from the 5' end, thereby supporting the interpretation that in the reverse transcriptase, the cleavage site for the RNase H is held at around this distance behind the DNA polymerase activity. In other words, it appears that for the intact protein, the RNase H and reverse transcriptase activities may work in a coupled or coordinate manner. We also found that more than 80% of the residual 5' oligonucleotides remained base paired to the RNA-directed DNA product. Furthermore, under certain conditions, these short RNAs could act as efficient primers for an associated DNA-directed DNA synthesis in the reverse direction.
J Virol. 1992 July; 66(7): 4271-4278
This article has been cited by other articles:
-
Duggal, N. K., Goo, L., King, S. R., Telesnitsky, A.
(2007). Effects of Identity Minimization on Moloney Murine Leukemia Virus Template Recognition and Frequent Tertiary Template-Directed Insertions during Nonhomologous Recombination. J. Virol.
81: 12156-12168
[Abstract]
[Full Text]
-
Schultz, S. J., Zhang, M., Champoux, J. J.
(2006). Sequence, Distance, and Accessibility Are Determinants of 5'-End-directed Cleavages by Retroviral RNases H. J. Biol. Chem.
281: 1943-1955
[Abstract]
[Full Text]
-
Miles, L. R., Agresta, B. E., Khan, M. B., Tang, S., Levin, J. G., Powell, M. D.
(2005). Effect of Polypurine Tract (PPT) Mutations on Human Immunodeficiency Virus Type 1 Replication: a Virus with a Completely Randomized PPT Retains Low Infectivity. J. Virol.
79: 6859-6867
[Abstract]
[Full Text]
-
Wisniewski, M., Chen, Y., Balakrishnan, M., Palaniappan, C., Roques, B. P., Fay, P. J., Bambara, R. A.
(2002). Substrate Requirements for Secondary Cleavage by HIV-1 Reverse Transcriptase RNase H. J. Biol. Chem.
277: 28400-28410
[Abstract]
[Full Text]
-
Julias, J. G., Ferris, A. L., Boyer, P. L., Hughes, S. H.
(2001). Replication of Phenotypically Mixed Human Immunodeficiency Virus Type 1 Virions Containing Catalytically Active and Catalytically Inactive Reverse Transcriptase. J. Virol.
75: 6537-6546
[Abstract]
[Full Text]
-
Wisniewski, M., Balakrishnan, M., Palaniappan, C., Fay, P. J., Bambara, R. A.
(2000). Unique progressive cleavage mechanism of HIV reverse transcriptase RNase H. Proc. Natl. Acad. Sci. USA
10.1073/pnas.210392297v1
[Abstract]
[Full Text]
-
Driscoll, M. D., Hughes, S. H.
(2000). Human Immunodeficiency Virus Type 1 Nucleocapsid Protein Can Prevent Self-Priming of Minus-Strand Strong Stop DNA by Promoting the Annealing of Short Oligonucleotides to Hairpin Sequences. J. Virol.
74: 8785-8792
[Abstract]
[Full Text]
-
Schultz, S. J., Zhang, M., Kelleher, C. D., Champoux, J. J.
(1999). Polypurine Tract Primer Generation and Utilization by Moloney Murine Leukemia Virus Reverse Transcriptase. J. Biol. Chem.
274: 34547-34555
[Abstract]
[Full Text]
-
Powell, M. D., Beard, W. A., Bebenek, K., Howard, K. J., Le Grice, S. F. J., Darden, T. A., Kunkel, T. A., Wilson, S. H., Levin, J. G.
(1999). Residues in the alpha H and alpha I Helices of the HIV-1 Reverse Transcriptase Thumb Subdomain Required for the Specificity of RNase H-catalyzed Removal of the Polypurine Tract Primer. J. Biol. Chem.
274: 19885-19893
[Abstract]
[Full Text]
-
Wu, T., Guo, J., Bess, J., Henderson, L. E., Levin, J. G.
(1999). Molecular Requirements for Human Immunodeficiency Virus Type 1 Plus-Strand Transfer: Analysis in Reconstituted and Endogenous Reverse Transcription Systems. J. Virol.
73: 4794-4805
[Abstract]
[Full Text]
-
Bowman, R. R., Hu, W.-S., Pathak, V. K.
(1998). Relative Rates of Retroviral Reverse Transcriptase Template Switching during RNA- and DNA-Dependent DNA Synthesis. J. Virol.
72: 5198-5206
[Abstract]
[Full Text]
-
Palaniappan, C., Wisniewski, M., Jacques, P. S., Le Grice, S. F.J., Fay, P. J., Bambara, R. A.
(1997). Mutations within the Primer Grip Region of HIV-1 Reverse Transcriptase Result in Loss of RNase H Function. J. Biol. Chem.
272: 11157-11164
[Abstract]
[Full Text]
-
DeStefano, J. J., DeStefano, J. J.
(1996). Interaction of Human Immunodeficiency Virus Nucleocapsid Protein with a Structure Mimicking a Replication Intermediate. EFFECTS ON STABILITY, REVERSE TRANSCRIPTASE BINDING, AND STRAND TRANSFER. J. Biol. Chem.
271: 16350-16356
[Abstract]
[Full Text]
-
Schultz, S. J., Whiting, S. H., Champoux, J. J.
(1995). Cleavage Specificities of Moloney Murine Leukemia Virus RNase H Implicated in the Second Strand Transfer During Reverse Transcription. J. Biol. Chem.
270: 24135-24145
[Abstract]
[Full Text]
-
Schultz, S. J., Zhang, M., Kelleher, C. D., Champoux, J. J.
(2000). Analysis of Plus-strand Primer Selection, Removal, and Reutilization by Retroviral Reverse Transcriptases. J. Biol. Chem.
275: 32299-32309
[Abstract]
[Full Text]
-
Wisniewski, M., Balakrishnan, M., Palaniappan, C., Fay, P. J., Bambara, R. A.
(2000). The Sequential Mechanism of HIV Reverse Transcriptase RNase H. J. Biol. Chem.
275: 37664-37671
[Abstract]
[Full Text]
-
DeStefano, J. J., Cristofaro, J. V., Derebail, S., Bohlayer, W. P., Fitzgerald-Heath, M. J.
(2001). Physical Mapping of HIV Reverse Transcriptase to the 5' End of RNA Primers. J. Biol. Chem.
276: 32515-32521
[Abstract]
[Full Text]
-
Wisniewski, M., Balakrishnan, M., Palaniappan, C., Fay, P. J., Bambara, R. A.
(2000). Unique progressive cleavage mechanism of HIV reverse transcriptase RNase H. Proc. Natl. Acad. Sci. USA
97: 11978-11983
[Abstract]
[Full Text]
Copyright © 1992 by the American Society for Microbiology. All rights reserved.