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Journal of Virology, April 2002, p. 4131-4137, Vol. 76, No. 8
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.8.4131-4137.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Role of RNA in Facilitating Gag/Gag-Pol Interaction

Ahmad Khorchid,1,2 Rabih Halwani,1,2 Mark A. Wainberg,1,2,3 and Lawrence Kleiman1,2,3*

Lady Davis Institute for Medical Research and McGill AIDS Center Jewish General Hospital,1 Departments of Medicine,2 Immunology and Microbiology, McGill University, Montreal, Quebec, Canada H3T 1E23

Received 17 September 2001/ Accepted 11 January 2002

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.

* Corresponding author. Mailing address: Lady Davis Institute for Medical Research-Jewish General Hospital, 3755 Cote Ste-Catherine Rd., Montreal, Quebec, Canada H3T 1E2. Phone: (514) 340-8260. Fax: (514) 340-7502. E-mail: lawrence.kleiman{at}mcgill.ca.

Journal of Virology, April 2002, p. 4131-4137, Vol. 76, No. 8
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.8.4131-4137.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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