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 Liang, C.
Right arrow Articles by Wainberg, M. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Liang, C.
Right arrow Articles by Wainberg, M. A.

 Previous Article  |  Next Article 

Journal of Virology, July 2000, p. 6251-6261, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Deletion Mutagenesis Downstream of the 5' Long Terminal Repeat of Human Immunodeficiency Virus Type 1 Is Compensated for by Point Mutations in both the U5 Region and gag Gene

Chen Liang,* Liwei Rong, Rodney S. Russell, and Mark A. Wainberg*

McGill AIDS Centre, Lady Davis Institute-Jewish General Hospital, Montreal, Québec, Canada H3T 1E2, and Department of Microbiology and Immunology, McGill University, Montreal, Québec, Canada H3A 2B4

Received 27 January 2000/Accepted 21 April 2000

We have studied the role of an RNA region at nucleotides (nt) +200 to +233, just downstream of the 5' long terminal repeat, in encapsidation of human immunodeficiency virus type 1 genomic RNA. Three deletion mutations, namely, BH-D0, BH-D1, and BH-D2, were generated to eliminate sequences at positions nt +200 to +219, +200 to +226, and +200 to +233. The result in each case was decreased levels of packaging of viral RNA into the mutated viruses, with the BH-D2 virus being the most severely affected. Consistently, all three deletions resulted in impaired viral infectiousness and the BH-D2 mutation showed the most dramatic impact in this regard. Further analysis revealed additional defects in Gag precursor processing and in the extension efficiency of the tRNA3Lys primer in reverse transcription reactions performed with these mutated viruses. To shed further light on the function of these deleted sequences in viral replication, the mutated viruses were cultured in MT-2 cells over prolonged periods to enable them to reacquire wild-type replication kinetics. Sequencing of the reverted viruses revealed point mutations in both the noncoding region and the gag gene. In the case of the BH-D0 revertant, two mutations were observed at positions G112A in the U5 region, termed M1, and T24I in the nucleocapsid protein, termed MNC, respectively. Either of these two mutations was able to confer wild-type replication capacity on BH-D0. In the case of BH-D1, each of the M1 mutations, a mutation termed M2, i.e., C227T, just downstream of the primer binding site, a mutation termed MP2 (T12I) in the p2 protein, and the MNC mutation were observed. A combination of either M1 and M2 or MP2 and MNC was able to rescue BH-D1. In the case of the BH-D2 deletion-containing viruses, three point mutations, i.e., M1, MP2, and MNC, were observed and the presence of all three was required to restore viral replication to wild-type levels.

* Corresponding author. Mailing address: McGill AIDS Centre, Lady Davis Institute-Jewish General Hospital, 3755 Cote Ste-Catherine Rd., Montreal, Québec, Canada H3T 1E2. Phone: (514) 340-8260. Fax: (514) 340-7537. E-mail: mdwa{at}musica.mcgill.ca.

Journal of Virology, July 2000, p. 6251-6261, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Dykes, C., Demeter, L. M. (2007). Clinical Significance of Human Immunodeficiency Virus Type 1 Replication Fitness. Clin. Microbiol. Rev. 20: 550-578 [Abstract] [Full Text]  
  • Russell, R. S., Roldan, A., Detorio, M., Hu, J., Wainberg, M. A., Liang, C. (2003). Effects of a Single Amino Acid Substitution within the p2 Region of Human Immunodeficiency Virus Type 1 on Packaging of Spliced Viral RNA. J. Virol. 77: 12986-12995 [Abstract] [Full Text]  
  • Guo, F., Cen, S., Niu, M., Javanbakht, H., Kleiman, L. (2003). Specific Inhibition of the Synthesis of Human Lysyl-tRNA Synthetase Results in Decreases in tRNALys Incorporation, tRNA3LysAnnealing to Viral RNA, and Viral Infectivity in Human Immunodeficiency Virus Type 1. J. Virol. 77: 9817-9822 [Abstract] [Full Text]  
  • Patel, J., Wang, S.-W., Izmailova, E., Aldovini, A. (2003). The Simian Immunodeficiency Virus 5' Untranslated Leader Sequence Plays a Role in Intracellular Viral Protein Accumulation and in RNA Packaging. J. Virol. 77: 6284-6292 [Abstract] [Full Text]  
  • Iwatani, Y., Rosen, A. E., Guo, J., Musier-Forsyth, K., Levin, J. G. (2003). Efficient Initiation of HIV-1 Reverse Transcription in Vitro. REQUIREMENT FOR RNA SEQUENCES DOWNSTREAM OF THE PRIMER BINDING SITE ABROGATED BY NUCLEOCAPSID PROTEIN-DEPENDENT PRIMER-TEMPLATE INTERACTIONS. J. Biol. Chem. 278: 14185-14195 [Abstract] [Full Text]  
  • Guan, Y., Diallo, K., Whitney, J. B., Liang, C., Wainberg, M. A. (2001). An Intact U5-Leader Stem Is Important for Efficient Replication of Simian Immunodeficiency Virus. J. Virol. 75: 11924-11929 [Abstract] [Full Text]  
  • Rong, L., Russell, R. S., Hu, J., Guan, Y., Kleiman, L., Liang, C., Wainberg, M. A. (2001). Hydrophobic Amino Acids in the Human Immunodeficiency Virus Type 1 p2 and Nucleocapsid Proteins Can Contribute to the Rescue of Deleted Viral RNA Packaging Signals. J. Virol. 75: 7230-7243 [Abstract] [Full Text]  
  • Guan, Y., Whitney, J. B., Liang, C., Wainberg, M. A. (2001). Novel, Live Attenuated Simian Immunodeficiency Virus Constructs Containing Major Deletions in Leader RNA Sequences. J. Virol. 75: 2776-2785 [Abstract] [Full Text]  
  • Smith, S. M., Khoroshev, M., Marx, P. A., Orenstein, J., Jeang, K.-T. (2001). Constitutively Dead, Conditionally Live HIV-1 Genomes. EX VIVO IMPLICATIONS FOR A LIVE VIRUS VACCINE. J. Biol. Chem. 276: 32184-32190 [Abstract] [Full Text]  
  • Beerens, N., Groot, F., Berkhout, B. (2001). Initiation of HIV-1 Reverse Transcription Is Regulated by a Primer Activation Signal. J. Biol. Chem. 276: 31247-31256 [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»