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 Driscoll, M. D.
Right arrow Articles by Hughes, S. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Driscoll, M. D.
Right arrow Articles by Hughes, S. H.

Next Article 

Journal of Virology, October 2000, p. 8785-8792, Vol. 74, No. 19
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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

Mark D. Driscolldagger and Stephen H. Hughes*

ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, Frederick, Maryland 21702-1201

Received 12 April 2000/Accepted 18 June 2000

Understanding how viral components collaborate to convert the human immunodeficiency virus type 1 genome from single-stranded RNA into double-stranded DNA is critical to the understanding of viral replication. Not only must the correct reactions be carried out, but unwanted side reactions must be avoided. After minus-strand strong stop DNA (-sssDNA) synthesis, degradation of the RNA template by the RNase H domain of reverse transcriptase (RT) produces single-stranded DNA that has the potential to self-prime at the imperfectly base-paired TAR hairpin, making continued DNA synthesis impossible. Although nucleocapsid protein (NC) interferes with -sssDNA self-priming in reverse transcription reactions in vitro, NC alone did not prevent self-priming of a synthetic -sssDNA oligomer. NC did not influence DNA bending and therefore cannot inhibit self-priming at hairpins by directly blocking hairpin formation. Using DNA oligomers as a model for genomic RNA fragments, we found that a 17-base DNA fragment annealed to the 3' end of the -sssDNA prevented self-priming in the presence of NC. This implies that to avoid self-priming, an RNA-DNA hybrid that is more thermodynamically stable than the hairpin must remain within the hairpin region. This suggests that NC prevents self-priming by generating or stabilizing a thermodynamically favored RNA-DNA heteroduplex instead of the kinetically favored TAR hairpin. In support of this idea, sequence changes that increased base pairing in the DNA TAR hairpin resulted in an increase in self-priming in vitro. We present a model describing the role of NC-dependent inhibition of self-priming in first-strand transfer.

* Corresponding author. Present address: HIV Drug Resistance Program, NCI-Frederick Cancer Research and Development Center, P.O. Box B, Frederick, MD 21702-1201. Phone: (301) 846-1619. Fax: (301) 846-6966. E-mail: hughes{at}ncifcrf.gov.

dagger Present address: HIV Drug Resistance Program, National Cancer Institute-FCRDC, Frederick, MD 21702-1201.

Journal of Virology, October 2000, p. 8785-8792, Vol. 74, No. 19
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Darugar, Q., Kim, H., Gorelick, R. J., Landes, C. (2008). Human T-Cell Lymphotropic Virus Type 1 Nucleocapsid Protein-Induced Structural Changes in Transactivation Response DNA Hairpin Measured by Single-Molecule Fluorescence Resonance Energy Transfer. J. Virol. 82: 12164-12171 [Abstract] [Full Text]  
  • Stewart-Maynard, K. M., Cruceanu, M., Wang, F., Vo, M.-N., Gorelick, R. J., Williams, M. C., Rouzina, I., Musier-Forsyth, K. (2008). Retroviral Nucleocapsid Proteins Display Nonequivalent Levels of Nucleic Acid Chaperone Activity. J. Virol. 82: 10129-10142 [Abstract] [Full Text]  
  • Song, M., Basu, V. P., Hanson, M. N., Roques, B. P., Bambara, R. A. (2008). Proximity and Branch Migration Mechanisms in HIV-1 Minus Strand Strong Stop DNA Transfer. J. Biol. Chem. 283: 3141-3150 [Abstract] [Full Text]  
  • Wu, T., Heilman-Miller, S. L., Levin, J. G. (2007). Effects of nucleic acid local structure and magnesium ions on minus-strand transfer mediated by the nucleic acid chaperone activity of HIV-1 nucleocapsid protein. Nucleic Acids Res 35: 3974-3987 [Abstract] [Full Text]  
  • Song, M., Balakrishnan, M., Chen, Y., Roques, B. P., Bambara, R. A. (2006). Stimulation of HIV-1 Minus Strand Strong Stop DNA Transfer by Genomic Sequences 3' of the Primer Binding Site. J. Biol. Chem. 281: 24227-24235 [Abstract] [Full Text]  
  • Cruceanu, M., Urbaneja, M. A., Hixson, C. V., Johnson, D. G., Datta, S. A., Fivash, M. J., Stephen, A. G., Fisher, R. J., Gorelick, R. J., Casas-Finet, J. R., Rein, A., Rouzina, I., Williams, M. C. (2006). Nucleic acid binding and chaperone properties of HIV-1 Gag and nucleocapsid proteins. Nucleic Acids Res 34: 593-605 [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, Y., Balakrishnan, M., Roques, B. P., Bambara, R. A. (2005). Acceptor RNA Cleavage Profile Supports an Invasion Mechanism for HIV-1 Minus Strand Transfer. J. Biol. Chem. 280: 14443-14452 [Abstract] [Full Text]  
  • Heilman-Miller, S. L., Wu, T., Levin, J. G. (2004). Alteration of Nucleic Acid Structure and Stability Modulates the Efficiency of Minus-Strand Transfer Mediated by the HIV-1 Nucleocapsid Protein. J. Biol. Chem. 279: 44154-44165 [Abstract] [Full Text]  
  • Chen, Y., Balakrishnan, M., Roques, B. P., Bambara, R. A. (2003). Steps of the Acceptor Invasion Mechanism for HIV-1 Minus Strand Strong Stop Transfer. J. Biol. Chem. 278: 38368-38375 [Abstract] [Full Text]  
  • Lee, N., Gorelick, R. J., Musier-Forsyth, K. (2003). Zinc finger-dependent HIV-1 nucleocapsid protein-TAR RNA interactions. Nucleic Acids Res 31: 4847-4855 [Abstract] [Full Text]  
  • Post, K., Guo, J., Howard, K. J., Powell, M. D., Miller, J. T., Hizi, A., Le Grice, S. F. J., Levin, J. G. (2003). Human Immunodeficiency Virus Type 2 Reverse Transcriptase Activity in Model Systems That Mimic Steps in Reverse Transcription. J. Virol. 77: 7623-7634 [Abstract] [Full Text]  
  • Derebail, S. S., Heath, M. J., DeStefano, J. J. (2003). Evidence for the Differential Effects of Nucleocapsid Protein on Strand Transfer in Various Regions of the HIV Genome. J. Biol. Chem. 278: 15702-15712 [Abstract] [Full Text]  
  • Chen, Y., Balakrishnan, M., Roques, B. P., Fay, P. J., Bambara, R. A. (2003). Mechanism of Minus Strand Strong Stop Transfer in HIV-1 Reverse Transcription. J. Biol. Chem. 278: 8006-8017 [Abstract] [Full Text]  
  • Fu, W., Hu, W.-S. (2002). Functional Replacement of Nucleocapsid Flanking Regions by Heterologous Counterparts with Divergent Primary Sequences: Effects of Chimeric Nucleocapsid on the Retroviral Replication Cycle. J. Virol. 77: 754-761 [Abstract] [Full Text]  
  • Roda, R. H., Balakrishnan, M., Kim, J. K., Roques, B. P., Fay, P. J., Bambara, R. A. (2002). Strand Transfer Occurs in Retroviruses by a Pause-initiated Two-step Mechanism. J. Biol. Chem. 277: 46900-46911 [Abstract] [Full Text]  
  • Zhang, W.-h., Hwang, C. K., Hu, W.-S., Gorelick, R. J., Pathak, V. K. (2002). Zinc Finger Domain of Murine Leukemia Virus Nucleocapsid Protein Enhances the Rate of Viral DNA Synthesis in Vivo. J. Virol. 76: 7473-7484 [Abstract] [Full Text]  
  • Guo, J., Wu, T., Kane, B. F., Johnson, D. G., Henderson, L. E., Gorelick, R. J., Levin, J. G. (2002). Subtle Alterations of the Native Zinc Finger Structures Have Dramatic Effects on the Nucleic Acid Chaperone Activity of Human Immunodeficiency Virus Type 1 Nucleocapsid Protein. J. Virol. 76: 4370-4378 [Abstract] [Full Text]  
  • Hameau, L., Jeusset, J., Lafosse, S., Coulaud, D., Delain, E., Unge, T., Restle, T., Le Cam, E., Mirambeau, G. (2001). Human Immunodeficiency Virus Type 1 Central DNA Flap: Dynamic Terminal Product of Plus-Strand Displacement DNA Synthesis Catalyzed by Reverse Transcriptase Assisted by Nucleocapsid Protein. J. Virol. 75: 3301-3313 [Abstract] [Full Text]  
  • Driscoll, M. D., Golinelli, M.-P., Hughes, S. H. (2001). In Vitro Analysis of Human Immunodeficiency Virus Type 1 Minus-Strand Strong-Stop DNA Synthesis and Genomic RNA Processing. J. Virol. 75: 672-686 [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»