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 Svarovskaia, E. S.
Right arrow Articles by Pathak, V. K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Svarovskaia, E. S.
Right arrow Articles by Pathak, V. K.

 Previous Article  |  Next Article 

Journal of Virology, April 2004, p. 3210-3222, Vol. 78, No. 7
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.7.3210-3222.2004

Azido-Containing Diketo Acid Derivatives Inhibit Human Immunodeficiency Virus Type 1 Integrase In Vivo and Influence the Frequency of Deletions at Two-Long-Terminal-Repeat-Circle Junctions

Evguenia S. Svarovskaia,1 Rebekah Barr,1 Xuechun Zhang,2 Godwin C. G. Pais,2 Christophe Marchand,3 Yves Pommier,3 Terrence R. Burke Jr.,2 and Vinay K. Pathak1*

HIV Drug Resistance Program,1 Laboratory of Medicinal Chemistry, Center for Cancer Research, National Cancer Institute at Frederick, Frederick, Maryland 21702,2 Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 208923

Received 8 May 2003/ Accepted 2 January 2004

We previously found that azido-containing ß-diketo acid derivatives (DKAs) are potent inhibitors of human immunodeficiency virus type 1 (HIV-1) integrase (IN) (X. Zhang et al., Bioorg. Med. Chem. Lett., 13:1215-1219, 2003). To characterize the intracellular mechanisms of action of DKAs, we analyzed the antiviral activities of two potent azido-containing DKAs with either a monosubstitution or a disubstitution of azido groups, using single- and multiple-replication-cycle assays. Both azido-containing DKAs significantly inhibited HIV-1 infection in 293T, CEM-SS, and H9 cells (50% inhibitory concentration = 2 to 13 µM) and exhibited low cytotoxicity (50% cytotoxic concentration = 60 to 600 µM). Inhibition of HIV-1 IN in vivo was demonstrated by the observation that previously described L-708,906 resistance mutations in HIV-1 IN (T66I and T66I/S153Y) also conferred resistance to the azido-group-containing DKAs. In vitro assays and in vivo analysis indicated that the DKAs did not significantly inhibit the 3' processing and selectively inhibited the strand transfer reaction. In addition, quantitative PCR indicated that two-long-terminal-repeat (2-LTR) circles were elevated in the presence of the azido-containing DKAs, confirming that HIV-1 IN was the intracellular target of viral inhibition. To gain insight into the mechanism by which the DKAs increased 2-LTR-circle formation of 3'-processed viral DNAs, we performed extensive DNA sequencing analysis of 2-LTR-circle junctions. The results indicated that the frequency of deletions at the circle junctions was elevated from 19% for the untreated controls to 32 to 41% in the presence of monosubstituted (but not disubstituted) DKAs. These results indicate that the structure of the DKAs can influence the extent of degradation of viral DNA ends by host nucleases and the frequency of deletions at the 2-LTR-circle junctions. Thus, sequencing analysis of 2-LTR-circle junctions can elucidate the intracellular mechanisms of action of HIV-1 IN inhibitors.

* Corresponding author. Mailing address: HIV Drug Resistance Program, National Cancer Institute—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, April 2004, p. 3210-3222, Vol. 78, No. 7
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.7.3210-3222.2004




This article has been cited by other articles:

  • Covens, K., Dekeersmaeker, N., Schrooten, Y., Weber, J., Schols, D., Quinones-Mateu, M. E., Vandamme, A.-M., Van Laethem, K. (2009). Novel Recombinant Virus Assay for Measuring Susceptibility of Human Immunodeficiency Virus Type 1 Group M Subtypes To Clinically Approved Drugs. J. Clin. Microbiol. 47: 2232-2242 [Abstract] [Full Text]  
  • Vatakis, D. N., Kim, S., Kim, N., Chow, S. A., Zack, J. A. (2009). Human Immunodeficiency Virus Integration Efficiency and Site Selection in Quiescent CD4+ T Cells. J. Virol. 83: 6222-6233 [Abstract] [Full Text]  
  • Vatakis, D. N., Nixon, C. C., Bristol, G., Zack, J. A. (2009). Differentially Stimulated CD4+ T Cells Display Altered Human Immunodeficiency Virus Infection Kinetics: Implications for the Efficacy of Antiviral Agents. J. Virol. 83: 3374-3378 [Abstract] [Full Text]  
  • Delelis, O., Malet, I., Na, L., Tchertanov, L., Calvez, V., Marcelin, A.-G., Subra, F., Deprez, E., Mouscadet, J.-F. (2009). The G140S mutation in HIV integrases from raltegravir-resistant patients rescues catalytic defect due to the resistance Q148H mutation. Nucleic Acids Res 37: 1193-1201 [Abstract] [Full Text]  
  • Bosque, A., Planelles, V. (2009). Induction of HIV-1 latency and reactivation in primary memory CD4+ T cells. Blood 113: 58-65 [Abstract] [Full Text]  
  • McKee, C. J., Kessl, J. J., Shkriabai, N., Dar, M. J., Engelman, A., Kvaratskhelia, M. (2008). Dynamic Modulation of HIV-1 Integrase Structure and Function by Cellular Lens Epithelium-derived Growth Factor (LEDGF) Protein. J. Biol. Chem. 283: 31802-31812 [Abstract] [Full Text]  
  • Ruggiero, E., Bona, R., Muratori, C., Federico, M. (2008). Virological Consequences of Early Events following Cell-Cell Contact between Human Immunodeficiency Virus Type 1-Infected and Uninfected CD4+ Cells. J. Virol. 82: 7773-7789 [Abstract] [Full Text]  
  • Pandey, K. K., Bera, S., Zahm, J., Vora, A., Stillmock, K., Hazuda, D., Grandgenett, D. P. (2007). Inhibition of Human Immunodeficiency Virus Type 1 Concerted Integration by Strand Transfer Inhibitors Which Recognize a Transient Structural Intermediate. J. Virol. 81: 12189-12199 [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]  
  • Cara, A., Klotman, M. E. (2006). Retroviral E-DNA: persistence and gene expression in nondividing immune cells. J. Leukoc. Biol. 80: 1013-1017 [Abstract] [Full Text]  
  • Bona, R., Andreotti, M., Buffa, V., Leone, P., Galluzzo, C. M., Amici, R., Palmisano, L., Mancini, M. G., Michelini, Z., Di Santo, R., Costi, R., Roux, A., Pommier, Y., Marchand, C., Vella, S., Cara, A. (2006). Development of a human immunodeficiency virus vector-based, single-cycle assay for evaluation of anti-integrase compounds.. Antimicrob. Agents Chemother. 50: 3407-3417 [Abstract] [Full Text]  
  • Mandal, D., Dash, C., Le Grice, S. F. J., Prasad, V. R. (2006). Analysis of HIV-1 replication block due to substitutions at F61 residue of reverse transcriptase reveals additional defects involving the RNase H function. Nucleic Acids Res 34: 2853-2863 [Abstract] [Full Text]  
  • Williams, K. L., Zhang, Y., Shkriabai, N., Karki, R. G., Nicklaus, M. C., Kotrikadze, N., Hess, S., Le Grice, S. F. J., Craigie, R., Pathak, V. K., Kvaratskhelia, M. (2005). Mass Spectrometric Analysis of the HIV-1 Integrase-Pyridoxal 5'-Phosphate Complex Reveals a New Binding Site for a Nucleotide Inhibitor. J. Biol. Chem. 280: 7949-7955 [Abstract] [Full Text]  
  • Svarovskaia, E. S., Xu, H., Mbisa, J. L., Barr, R., Gorelick, R. J., Ono, A., Freed, E. O., Hu, W.-S., Pathak, V. K. (2004). Human Apolipoprotein B mRNA-editing Enzyme-catalytic Polypeptide-like 3G (APOBEC3G) Is Incorporated into HIV-1 Virions through Interactions with Viral and Nonviral RNAs. J. Biol. Chem. 279: 35822-35828 [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»