Role of RNA in facilitating Gag/Gag-Pol interaction

Journal of Virology
Ahmad KhorchidLawrence Kleiman

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.

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Citations

Jan 26, 2006·Nucleic Acids Research·Robert J FisherAlan Rein
Mar 30, 2007·Journal of Virology·Samuel J RulliAlan Rein
Jan 8, 2011·Retrovirology·Jing MaShan Cen
Dec 3, 2010·PLoS Pathogens·Sebla B Kutluay, Paul D Bieniasz
Nov 17, 2009·FEBS Letters·Lawrence KleimanKarin Musier-Forsyth
May 7, 2013·Virus Research·Klaus B Andersen
Jun 16, 2012·Virus Research·Jenan Saadatmand, Lawrence Kleiman
Nov 14, 2006·Journal of Molecular Biology·Siddhartha A K DattaAlan Rein
Jul 19, 2014·Virus Research·Valerie Le SageFernando Valiente-Echeverría
Jul 11, 2014·Frontiers in Microbiology·José O MaldonadoLouis M Mansky
Feb 8, 2008·Virus Research·Truus E M Abbink, Ben Berkhout
Jul 26, 2012·Molecular Biology International·Ian B HogueAkira Ono
May 26, 2004·The Journal of Biological Chemistry·Shan CenLawrence Kleiman
May 13, 2005·The Journal of Biological Chemistry·Fei GuoLawrence Kleiman
Sep 26, 2020·Journal of Proteome Research·Jesús Lavado-GarcíaFrancesc Gòdia

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