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 Nikolenko, G. N.
Right arrow Articles by Pathak, V. K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nikolenko, G. N.
Right arrow Articles by Pathak, V. K.

 Previous Article  |  Next Article 

Journal of Virology, August 2004, p. 8761-8770, Vol. 78, No. 16
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.16.8761-8770.2004

Antiretroviral Drug Resistance Mutations in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Increase Template-Switching Frequency

Galina N. Nikolenko,1 Evguenia S. Svarovskaia,1 Krista A. Delviks,1 and Vinay K. Pathak1*

HIV Drug Resistance Program, National Cancer Institute at Frederick, Frederick, Maryland 217021

Received 2 February 2004/ Accepted 12 April 2004

Template-switching events during reverse transcription are necessary for completion of retroviral replication and recombination. Structural determinants of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) that influence its template-switching frequency are not known. To identify determinants of HIV-1 RT that affect the frequency of template switching, we developed an in vivo assay in which RT template-switching events during viral replication resulted in functional reconstitution of the green fluorescent protein gene. A survey of single amino acid substitutions near the polymerase active site or deoxynucleoside triphosphate-binding site of HIV-1 RT indicated that several substitutions increased the rate of RT template switching. Several mutations associated with resistance to antiviral nucleoside analogs (K65R, L74V, E89G, Q151N, and M184I) dramatically increased RT template-switching frequencies by two- to sixfold in a single replication cycle. In contrast, substitutions in the RNase H domain (H539N, D549N) decreased the frequency of RT template switching by twofold. Depletion of intracellular nucleotide pools by hydroxyurea treatment of cells used as targets for infection resulted in a 1.8-fold increase in the frequency of RT template switching. These results indicate that the dynamic steady state between polymerase and RNase H activities is an important determinant of HIV-1 RT template switching and establish that HIV-1 recombination occurs by the previously described dynamic copy choice mechanism. These results also indicate that mutations conferring resistance to antiviral drugs can increase the frequency of RT template switching and may influence the rate of retroviral recombination and viral evolution.

* Corresponding author. Mailing address: HIV Drug Resistance Program, NCI-Frederick, Bldg. 535, Rm. 334, Frederick, MD 21702. Phone: (301) 846-1710. Fax: (301) 846-6013. E-mail: VPATHAK{at}ncifcrf.gov.

Journal of Virology, August 2004, p. 8761-8770, Vol. 78, No. 16
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.16.8761-8770.2004




This article has been cited by other articles:

  • Roquebert, B., Marcelin, A.-G. (2008). The involvement of HIV-1 RNAse H in resistance to nucleoside analogues. J Antimicrob Chemother 61: 973-975 [Abstract] [Full Text]  
  • Mulder, L. C. F., Harari, A., Simon, V. (2008). Cytidine deamination induced HIV-1 drug resistance. Proc. Natl. Acad. Sci. USA 105: 5501-5506 [Abstract] [Full Text]  
  • Gao, L., Hanson, M. N., Balakrishnan, M., Boyer, P. L., Roques, B. P., Hughes, S. H., Kim, B., Bambara, R. A. (2008). Apparent Defects in Processive DNA Synthesis, Strand Transfer, and Primer Elongation of Met-184 Mutants of HIV-1 Reverse Transcriptase Derive Solely from a dNTP Utilization Defect. J. Biol. Chem. 283: 9196-9205 [Abstract] [Full Text]  
  • Hachiya, A., Kodama, E. N., Sarafianos, S. G., Schuckmann, M. M., Sakagami, Y., Matsuoka, M., Takiguchi, M., Gatanaga, H., Oka, S. (2008). Amino Acid Mutation N348I in the Connection Subdomain of Human Immunodeficiency Virus Type 1 Reverse Transcriptase Confers Multiclass Resistance to Nucleoside and Nonnucleoside Reverse Transcriptase Inhibitors. J. Virol. 82: 3261-3270 [Abstract] [Full Text]  
  • King, S. R., Duggal, N. K., Ndongmo, C. B., Pacut, C., Telesnitsky, A. (2008). Pseudodiploid Genome Organization Aids Full-Length Human Immunodeficiency Virus Type 1 DNA Synthesis. J. Virol. 82: 2376-2384 [Abstract] [Full Text]  
  • Ntemgwa, M., Wainberg, M. A., Oliveira, M., Moisi, D., Lalonde, R., Micheli, V., Brenner, B. G. (2007). Variations in Reverse Transcriptase and RNase H Domain Mutations in Human Immunodeficiency Virus Type 1 Clinical Isolates Are Associated with Divergent Phenotypic Resistance to Zidovudine. Antimicrob. Agents Chemother. 51: 3861-3869 [Abstract] [Full Text]  
  • Nora, T., Charpentier, C., Tenaillon, O., Hoede, C., Clavel, F., Hance, A. J. (2007). Contribution of Recombination to the Evolution of Human Immunodeficiency Viruses Expressing Resistance to Antiretroviral Treatment. J. Virol. 81: 7620-7628 [Abstract] [Full Text]  
  • Delviks-Frankenberry, K. A., Nikolenko, G. N., Barr, R., Pathak, V. K. (2007). Mutations in Human Immunodeficiency Virus Type 1 RNase H Primer Grip Enhance 3'-Azido-3'-Deoxythymidine Resistance. J. Virol. 81: 6837-6845 [Abstract] [Full Text]  
  • Mbisa, J. L., Barr, R., Thomas, J. A., Vandegraaff, N., Dorweiler, I. J., Svarovskaia, E. S., Brown, W. L., Mansky, L. M., Gorelick, R. J., Harris, R. S., Engelman, A., Pathak, V. K. (2007). Human Immunodeficiency Virus Type 1 cDNAs Produced in the Presence of APOBEC3G Exhibit Defects in Plus-Strand DNA Transfer and Integration. J. Virol. 81: 7099-7110 [Abstract] [Full Text]  
  • Villena, C., Prado, J. G., Puertas, M. C., Martinez, M. A., Clotet, B., Ruiz, L., Parkin, N. T., Menendez-Arias, L., Martinez-Picado, J. (2007). Relative Fitness and Replication Capacity of a Multinucleoside Analogue-Resistant Clinical Human Immunodeficiency Virus Type 1 Isolate with a Deletion of Codon 69 in the Reverse Transcriptase Coding Region. J. Virol. 81: 4713-4721 [Abstract] [Full Text]  
  • Nikolenko, G. N., Delviks-Frankenberry, K. A., Palmer, S., Maldarelli, F., Fivash, M. J. Jr., Coffin, J. M., Pathak, V. K. (2007). Mutations in the connection domain of HIV-1 reverse transcriptase increase 3'-azido-3'-deoxythymidine resistance. Proc. Natl. Acad. Sci. USA 104: 317-322 [Abstract] [Full Text]  
  • Operario, D. J., Balakrishnan, M., Bambara, R. A., Kim, B. (2006). Reduced dNTP Interaction of Human Immunodeficiency Virus Type 1 Reverse Transcriptase Promotes Strand Transfer. J. Biol. Chem. 281: 32113-32121 [Abstract] [Full Text]  
  • Charpentier, C., Nora, T., Tenaillon, O., Clavel, F., Hance, A. J. (2006). Extensive Recombination among Human Immunodeficiency Virus Type 1 Quasispecies Makes an Important Contribution to Viral Diversity in Individual Patients. J. Virol. 80: 2472-2482 [Abstract] [Full Text]  
  • Purohit, V., Balakrishnan, M., Kim, B., Bambara, R. A. (2005). Evidence That HIV-1 Reverse Transcriptase Employs the DNA 3' End-directed Primary/Secondary RNase H Cleavage Mechanism during Synthesis and Strand Transfer. J. Biol. Chem. 280: 40534-40543 [Abstract] [Full Text]  
  • Chen, J., Rhodes, T. D., Hu, W.-S. (2005). Comparison of the Genetic Recombination Rates of Human Immunodeficiency Virus Type 1 in Macrophages and T Cells. J. Virol. 79: 9337-9340 [Abstract] [Full Text]  
  • Nikolenko, G. N., Palmer, S., Maldarelli, F., Mellors, J. W., Coffin, J. M., Pathak, V. K. (2005). Mechanism for nucleoside analog-mediated abrogation of HIV-1 replication: Balance between RNase H activity and nucleotide excision. Proc. Natl. Acad. Sci. USA 102: 2093-2098 [Abstract] [Full Text]  
  • Mbisa, J. L., Nikolenko, G. N., Pathak, V. K. (2005). Mutations in the RNase H Primer Grip Domain of Murine Leukemia Virus Reverse Transcriptase Decrease Efficiency and Accuracy of Plus-Strand DNA Transfer. J. Virol. 79: 419-427 [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»