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A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
Published on: August 31, 2014
Evasion of CARD8 Activation During HIV-1 Assembly
Ivy K Hughes1, James B Hood1, Andrés A Quiñones-Molina1
1Department of Virology, Immunology, & Microbiology, Boston University Chobanian & Avedisian School of Medicine, Boston, MA 02118.
Abstract:
As intracellular parasites, viruses must devise sophisticated mechanisms to produce and assemble viral components while suppressing activation of innate immune effectors. Here, we report that coordination of HIV-1 assembly by the viral polyprotein Gag suppresses inappropriately-timed protease (PR) activity to evade the PR activity sensor, CARD8. Employing mutants of Gag, we show that disruption of domains controlling viral assembly site (MA) or virus particle release (NC and p6) lead to premature activation of PR and the CARD8 inflammasome, resulting in IL-1β secretion and pyroptotic cell death. Further, we demonstrate that previously-observed host-adaptive mutations in HIV-1 MA (M30K) and p6 (PTAP duplication) associated with greater fitness in humans differentially modulate the process of viral assembly and budding to evade CARD8-mediated cell death. Altogether, this work reveals adaptation to human CARD8 by HIV-1 Gag upon zoonotic transmission from chimpanzees and suggests that assembly-regulated CARD8 activation influences the trajectory of HIV-1 evolution and fitness in humans.
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