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 Zhang, W.-h.
Right arrow Articles by Pathak, V. K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zhang, W.-h.
Right arrow Articles by Pathak, V. K.

 Previous Article  |  Next Article 

Journal of Virology, August 2002, p. 7473-7484, Vol. 76, No. 15
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.15.7473-7484.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Zinc Finger Domain of Murine Leukemia Virus Nucleocapsid Protein Enhances the Rate of Viral DNA Synthesis in Vivo

Wen-hui Zhang,1,2 Carey K. Hwang,1,3 Wei-Shau Hu,1 Robert J. Gorelick,4 and Vinay K. Pathak1*

HIV Drug Resistance Program,1 AIDS Vaccine Program, Science Applications International Corporation Frederick, National Cancer Institute at Frederick, Frederick, Maryland 21702,4 Department of Biology,2 Department of Microbiology and Immunology, West Virginia University, Morgantown, West Virginia 265063

Received 14 November 2001/ Accepted 22 April 2002

In vitro studies have indicated that retroviral nucleocapsid (NC) protein facilitates both DNA synthesis by reverse transcriptase (RT) and annealing of the nascent DNA with acceptor template. Increasing the rate of DNA synthesis is expected to reduce the frequency of RT template switching, whereas annealing the nascent DNA with acceptor template promotes template switching. We performed a mutational analysis of the murine leukemia virus (MLV) NC zinc finger domain to study its effect on RT template switching in vivo and to explore the role of NC during reverse transcription. The effects of NC mutations on RT template switching were determined by using a previously described in vivo direct-repeat deletion assay. A trans-complementation assay was also developed in which replication-defective NC mutants were rescued by coexpression of replication-defective RT mutants that provided wild-type NC in trans. We found that mutations in the MLV NC zinc finger domain increased the frequency of template switching approximately twofold. When a predicted stem-loop RNA secondary structure was introduced into the template RNA, the template-switching frequency increased 5-fold for wild-type NC and further increased up to an additional 6-fold for NC zinc finger domain mutants, resulting in an overall increase of as much as 30-fold. Thus, wild-type NC increased the efficiency with which RT was able to reverse transcribe through regions of RNA secondary structure that might serve as RT pause sites. These results provide the first in vivo evidence that NC enhances the rate of DNA synthesis by RT in regions of the template possessing stable RNA secondary structure.

* 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, August 2002, p. 7473-7484, Vol. 76, No. 15
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.15.7473-7484.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Onafuwa-Nuga, A., Telesnitsky, A. (2009). The Remarkable Frequency of Human Immunodeficiency Virus Type 1 Genetic Recombination. Microbiol. Mol. Biol. Rev. 73: 451-480 [Abstract] [Full Text]  
  • Larsen, L. S. Z., Beliakova-Bethell, N., Bilanchone, V., Zhang, M., Lamsa, A., DaSilva, R., Hatfield, G. W., Nagashima, K., Sandmeyer, S. (2008). Ty3 Nucleocapsid Controls Localization of Particle Assembly. J. Virol. 82: 2501-2514 [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]  
  • Purohit, V., Roques, B. P., Kim, B., Bambara, R. A. (2007). Mechanisms That Prevent Template Inactivation by HIV-1 Reverse Transcriptase RNase H Cleavages. J. Biol. Chem. 282: 12598-12609 [Abstract] [Full Text]  
  • Gao, L., Balakrishnan, M., Roques, B. P., Bambara, R. A. (2007). Insights into the Multiple Roles of Pausing in HIV-1 Reverse Transcriptase-promoted Strand Transfers. J. Biol. Chem. 282: 6222-6231 [Abstract] [Full Text]  
  • Lanciault, C., Champoux, J. J. (2006). Pausing during Reverse Transcription Increases the Rate of Retroviral Recombination. J. Virol. 80: 2483-2494 [Abstract] [Full Text]  
  • Alamgir, A. S. M., Owens, N., Lavignon, M., Malik, F., Evans, L. H. (2005). Precise Identification of Endogenous Proviruses of NFS/N Mice Participating in Recombination with Moloney Ecotropic Murine Leukemia Virus (MuLV) To Generate Polytropic MuLVs. J. Virol. 79: 4664-4671 [Abstract] [Full Text]  
  • Lee, S.-K., Nagashima, K., Hu, W.-S. (2005). Cooperative Effect of Gag Proteins p12 and Capsid during Early Events of Murine Leukemia Virus Replication. J. Virol. 79: 4159-4169 [Abstract] [Full Text]  
  • Lanciault, C., Champoux, J. J. (2005). Effects of Unpaired Nucleotides within HIV-1 Genomic Secondary Structures on Pausing and Strand Transfer. J. Biol. Chem. 280: 2413-2423 [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]  
  • 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]  
  • Voronin, Y. A., Pathak, V. K. (2004). Frequent Dual Initiation in Human Immunodeficiency Virus-Based Vectors Containing Two Primer-Binding Sites: a Quantitative In Vivo Assay for Function of Initiation Complexes. J. Virol. 78: 5402-5413 [Abstract] [Full Text]  
  • Chen, M.-H., Icenogle, J. P. (2004). Rubella Virus Capsid Protein Modulates Viral Genome Replication and Virus Infectivity. J. Virol. 78: 4314-4322 [Abstract] [Full Text]  
  • Cheslock, S. R., Poon, D. T. K., Fu, W., Rhodes, T. D., Henderson, L. E., Nagashima, K., McGrath, C. F., Hu, W.-S. (2003). Charged Assembly Helix Motif in Murine Leukemia Virus Capsid: an Important Region for Virus Assembly and Particle Size Determination. J. Virol. 77: 7058-7066 [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]  


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