This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Drosopoulos, W. C.
Right arrow Articles by Prasad, V. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Drosopoulos, W. C.
Right arrow Articles by Prasad, V. R.

 Previous Article  |  Next Article 

J Virol, May 1998, p. 4224-4230, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Increased Misincorporation Fidelity Observed for Nucleoside Analog Resistance Mutations M184V and E89G in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Does Not Correlate with the Overall Error Rate Measured In Vitro

William C. Drosopoulos and Vinayaka R. Prasad*

Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York 10461

Received 9 December 1997/Accepted 29 January 1998

Nucleoside analog-resistant variants of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) that displayed higher in vitro polymerase fidelity were previously identified via nucleotide insertion and mispair extension assays. To evaluate the contribution of increased nucleotide insertion and primer extension fidelities on the overall error rate of HIV-1 RT, we have measured the impact of two such mutations, E89G and M184V, on DNA copying fidelity in an M13 phage-based forward mutation assay. Using this assay, we observed mutation frequencies of 8.60 × 10-3, 6.26 × 10-3, 5.53 × 10-3, and 12.30 × 10-3 for wild-type, E89G, M184V, and double-mutant E89G/M184V HIV-1 RTs, respectively. Therefore, the overall polymerase fidelities of wild-type, E89G, M184V, and E89G/M184V HIV-1 RTs are similar (less than twofold differences) for DNA-dependent DNA synthesis. Thus, rather large increases in fidelity of deoxynucleoside triphosphate insertion and mispair extension observed previously appear not to influence the overall error rate of these mutants. However, a qualitative analysis of the mutations induced revealed significant differences in the mutational spectra between the wild-type and mutant enzymes.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461. Phone: (718) 430-2517. Fax: (718) 430-8976. E-mail: prasad{at}aecom.yu.edu.

J Virol, May 1998, p. 4224-4230, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Hamburgh, M. E., Curr, K. A., Monaghan, M., Rao, V. R., Tripathi, S., Preston, B. D., Sarafianos, S., Arnold, E., Darden, T., Prasad, V. R. (2006). Structural Determinants of Slippage-mediated Mutations by Human Immunodeficiency Virus Type 1 Reverse Transcriptase. J. Biol. Chem. 281: 7421-7428 [Abstract] [Full Text]  
  • Curr, K., Tripathi, S., Lennerstrand, J., Larder, B. A., Prasad, V. R. (2006). Influence of naturally occurring insertions in the fingers subdomain of human immunodeficiency virus type 1 reverse transcriptase on polymerase fidelity and mutation frequencies in vitro. J. Gen. Virol. 87: 419-428 [Abstract] [Full Text]  
  • Skasko, M., Weiss, K. K., Reynolds, H. M., Jamburuthugoda, V., Lee, K., Kim, B. (2005). Mechanistic Differences in RNA-dependent DNA Polymerization and Fidelity between Murine Leukemia Virus and HIV-1 Reverse Transcriptases. J. Biol. Chem. 280: 12190-12200 [Abstract] [Full Text]  
  • Nikolenko, G. N., Svarovskaia, E. S., Delviks, K. A., Pathak, V. K. (2004). Antiretroviral Drug Resistance Mutations in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Increase Template-Switching Frequency. J. Virol. 78: 8761-8770 [Abstract] [Full Text]  
  • Sharma, B., Kaushik, N., Upadhyay, A., Tripathi, S., Singh, K., Pandey, V. N. (2003). A positively charged side chain at position 154 on the {beta}8-{alpha}E loop of HIV-1 RT is required for stable ternary complex formation. Nucleic Acids Res 31: 5167-5174 [Abstract] [Full Text]  
  • Pfeiffer, J. K., Kirkegaard, K. (2003). A single mutation in poliovirus RNA-dependent RNA polymerase confers resistance to mutagenic nucleotide analogs via increased fidelity. Proc. Natl. Acad. Sci. USA 100: 7289-7294 [Abstract] [Full Text]  
  • Fisher, T. S., Darden, T., Prasad, V. R. (2003). Mutations Proximal to the Minor Groove-Binding Track of Human Immunodeficiency Virus Type 1 Reverse Transcriptase Differentially Affect Utilization of RNA versus DNA as Template. J. Virol. 77: 5837-5845 [Abstract] [Full Text]  
  • Mansky, L. M., Le Rouzic, E., Benichou, S., Gajary, L. C. (2003). Influence of Reverse Transcriptase Variants, Drugs, and Vpr on Human Immunodeficiency Virus Type 1 Mutant Frequencies. J. Virol. 77: 2071-2080 [Abstract] [Full Text]  
  • Murry, J. P., Higgins, J., Matthews, T. B., Huang, V. Y., Van Rompay, K. K. A., Pedersen, N. C., North, T. W. (2002). Reversion of the M184V Mutation in Simian Immunodeficiency Virus Reverse Transcriptase Is Selected by Tenofovir, Even in the Presence of Lamivudine. J. Virol. 77: 1120-1130 [Abstract] [Full Text]  
  • Zhang, W.-H., Svarovskaia, E. S., Barr, R., Pathak, V. K. (2002). Y586F mutation in murine leukemia virus reverse transcriptase decreases fidelity of DNA synthesis in regions associated with adenine-thymine tracts. Proc. Natl. Acad. Sci. USA 99: 10090-10095 [Abstract] [Full Text]  
  • Fisher, T. S., Prasad, V. R. (2002). Substitutions of Phe61 Located in the Vicinity of Template 5'-Overhang Influence Polymerase Fidelity and Nucleoside Analog Sensitivity of HIV-1 Reverse Transcriptase. J. Biol. Chem. 277: 22345-22352 [Abstract] [Full Text]  
  • Shafer, R. W. (2002). Genotypic Testing for Human Immunodeficiency Virus Type 1 Drug Resistance. Clin. Microbiol. Rev. 15: 247-277 [Abstract] [Full Text]  
  • Gutierrez-Rivas, M., Menendez-Arias, L. (2001). A mutation in the primer grip region of HIV-1 reverse transcriptase that confers reduced fidelity of DNA synthesis. Nucleic Acids Res 29: 4963-4972 [Abstract] [Full Text]  
  • Mansky, L. M., Bernard, L. C. (2000). 3'-Azido-3'-Deoxythymidine (AZT) and AZT-Resistant Reverse Transcriptase Can Increase the In Vivo Mutation Rate of Human Immunodeficiency Virus Type 1. J. Virol. 74: 9532-9539 [Abstract] [Full Text]  
  • Boyer, P. L., Hughes, S. H. (2000). Effects of Amino Acid Substitutions at Position 115 on the Fidelity of Human Immunodeficiency Virus Type 1 Reverse Transcriptase. J. Virol. 74: 6494-6500 [Abstract] [Full Text]  
  • Halvas, E. K., Svarovskaia, E. S., Pathak, V. K. (2000). Development of an In Vivo Assay To Identify Structural Determinants in Murine Leukemia Virus Reverse Transcriptase Important for Fidelity. J. Virol. 74: 312-319 [Abstract] [Full Text]  
  • Wisniewski, M., Palaniappan, C., Fu, Z., Le Grice, S. F. J., Fay, P., Bambara, R. A. (1999). Mutations in the Primer Grip Region of HIV Reverse Transcriptase Can Increase Replication Fidelity. J. Biol. Chem. 274: 28175-28184 [Abstract] [Full Text]  
  • Shah, F. S., Curr, K. A., Hamburgh, M. E., Parniak, M., Mitsuya, H., Arnez, J. G., Prasad, V. R. (2000). Differential Influence of Nucleoside Analog-resistance Mutations K65R and L74V on the Overall Mutation Rate and Error Specificity of Human Immunodeficiency Virus Type 1 Reverse Transcriptase. J. Biol. Chem. 275: 27037-27044 [Abstract] [Full Text]  
  • Diamond, T. L., Kimata, J., Kim, B. (2001). Identification of a Simian Immunodeficiency Virus Reverse Transcriptase Variant with Enhanced Replicational Fidelity in the Late Stage of Viral Infection. J. Biol. Chem. 276: 23624-23631 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»