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PMID: 11907255 Published · ppublish English Journal Article

Role of RNA in facilitating Gag/Gag-Pol interaction.

Journal of virology ·Vol. 76 ·No. 8 ·2002-04-00 ·Pages 4131-7

Khorchid A, Halwani R, Wainberg MA, Kleiman L

Abstract

We have examined the influence of RNA upon the interaction of Gag-Pol with Gag during human immunodeficiency virus type 1 (HIV-1) assembly. COS7 cells were transfected with protease-negative HIV-1 proviral DNA, and Gag/Gag-Pol complexes were detected by coimmunoprecipitation with anti-integrase. In COS7 cells, Gag/Gag-Pol is found almost entirely in pelletable, membrane-bound complexes. Exposure of cells to 1% Triton X-100 releases Gag/Gag-Pol from bulk membrane, but the complexes remain pelletable. The role of RNA in facilitating the interaction between Gag and Gag-Pol was examined in these bulk membrane-free, pelletable complexes. The specific presence of viral genomic RNA is not required to maintain the Gag/Gag-Pol interaction, but some type of RNA is, since exposure to RNase destabilized the Gag/Gag-Pol complex. When present only in Gag, the nucleocapsid mutation R7R10K11S, which inhibits Gag binding to RNA, inhibits the formation of both Gag and Gag/Gag-Pol complexes. When present only in Gag-Pol, this mutation has no effect upon complex formation. This result indicates that Gag-Pol may not interact directly with RNA but rather requires RNA-facilitated Gag multimerization for its interaction with Gag.

MeSH Terms
Animals COS Cells Chlorocebus aethiops DNA, Viral/genetics,metabolism Fusion Proteins, gag-pol/metabolism Gene Products, gag/metabolism HIV-1/genetics,metabolism Humans Proviruses RNA, Viral/metabolism Transfection Virus Assembly
Chemicals
DNA, Viral Fusion Proteins, gag-pol Gene Products, gag RNA, Viral
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Khorchid Ahmad
Lady Davis Institute for Medical Research and McGill AIDS Center Jewish General Hospital, McGill University, Montreal, Quebec, Canada H3T 1E2.
Halwani Rabih
Wainberg Mark A
Kleiman Lawrence
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.76.8.4131-4137.2002
Published
2002-04-00
Pages
4131-7
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC136082
Subset
IM
External Links
PubMed source DOI: 10.1128/jvi.76.8.4131-4137.2002 PMC Full Text View journal details
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