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Abstract
GAGPOL DOMAINS RESPONSIBLE FOR THE LOCALIZATION OF GAG/GAGPOL COMPLEX TO DRM MUCH FASTER THAN GAG/GAG COMPLEXES
R. Halwani1-2, M.A. Wainberg1-2-3, L. Kleiman1-2-3
1Lady Davis Institute for Medical Research and McGill AIDS Centre, Jewish General Hospital, 2Departments of Medicine, 3Immunology and Microbiology, McGill University, Montreal Quebec, H3T 1E2 Canada
The purpose of this work is to study the movement of Gag/GagPol complex to the different membrane fractions during HIV-1 assembly, and to determine the Pol domains responsible of such movement. We have shown that in HIV-1-transfected COS7 cells, almost all steady-state Gag/Gag and Gag/GagPol complexes are membrane-bound. However, exposure to 1% Triton X-100 gives results indicating that while all Gag/GagPol complexes remain associated with detergent-resistant membrane (DRM), only 30% of Gag/Gag complexes are associated with DRM. Analysis of the localization of newly-synthesized Gag/Gag and Gag/GagPol to membrane indicates that after a 10 minute pulse with radioactive 35S-Cys/Met, all newly-synthesized Gag/GagPol is found at the DRM. Only 30% of newly-synthesized Gag/Gag moves to the membrane, and at 0 minute chase, only 38% of this membrane-bound Gag/Gag is associated with DRM. During the first 30 minutes of chase, most membrane-bound Gag/Gag moves to the DRM, while between 30 and 60 minutes chase, there is a significant decrease in membrane-bound Gag/Gag and Gag/GagPol. Since the localization of newly-synthesized Gag/Gag to DRM and the interaction of GagPol with Gag both depend upon Gag multimerization, the rapid localization of GagPol to DRM probably reflects the interaction of all newly-synthesized GagPol with the first new-synthesized polymeric Gag to associate with the DRM. We are now examining the domains of GagPol responsible of its rapid localization to DRM. Our preliminary data shows that regions upstream of INT within Pol domain are the one responsible of such movement.
The 2nd IAS Conference on HIV Pathogenesis and Treatment
Abstract no.
190
Suggested Citation
" R. Halwani -2, et al.
GAGPOL DOMAINS RESPONSIBLE FOR THE LOCALIZATION OF GAG/GAGPOL COMPLEX TO DRM MUCH FASTER THAN GAG/GAG COMPLEXES.
Oral Abstract:
The 2nd IAS Conference on HIV Pathogenesis and Treatment:
Abstract no.
190"
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