biostudies-literatureHughes IKNIDA NIH HHSNIAID NIH HHSeadz1069https://www.ebi.ac.uk/biostudies/studies/S-EPMC12609157biostudies-literatureUnknown11(46)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, caspase recruitment domain-containing protein 8 (CARD8). Using mutants of Gag, we show that disruption of domains controlling viral assembly site [matrix (MA)] or virus particle release (nucleocapsid and p6) leads to premature activation of PR and the CARD8 inflammasome, resulting in interleukin-1β (IL-1β) secretion and pyroptotic cell death. Further, we demonstrate that previously observed host-adaptive mutations in HIV-1 MA (M30K) and p6 (PTAP duplicatiScience advancesEvasion of CARD8 activation during HIV-1 assembly.PMC12609157R01 AI187175R01 DA051889R01 DA059952P30 AI042853R01 DA055488Hughes IKAkiyama HHood JBGummuluru SQuinones-Molina AAfalseEvasion of CARD8 activation during HIV-1 assembly.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, caspase recruitment domain-containing protein 8 (CARD8). Using mutants of Gag, we show that disruption of domains controlling viral assembly site [matrix (MA)] or virus particle release (nucleocapsid and p6) leads to premature activation of PR and the CARD8 inflammasome, resulting in interleukin-1β (IL-1β) secretion and pyroptotic cell death. Further, we demonstrate that previously observed host-adaptive mutations in HIV-1 MA (M30K) and p6 (PTAP duplicati2025-01-01T00:00:00Z2025 Nov2026-06-05T16:14:14.89Z2026-05-18T03:12:51.476ZS-EPMC126091574122328210.1126/sciadv.adz1069