In vivo HIV-1 nuclear condensates safeguard against cGAS and license reverse transcription

Entry of viral capsids into the nucleus induces the formation of biomolecular condensates called HIV-1 membraneless organelles (HIV-1-MLOs). Several questions remain about their persistence, in vivo formation, composition, and function. Our study reveals that HIV-1-MLOs persisted for several weeks in infected cells, and their abundance correlated with viral infectivity. Using an appropriate animal model, we show that HIV-1-MLOs were formed in vivo during acute infection. To explore the viral structures present within these biomolecular condensates, we used a combination of double immunogold labeling, electron microscopy and tomography, and unveiled a diverse array of viral core structures. Our functional analyses showed that HIV-1-MLOs remained stable during treatment with a reverse transcriptase inhibitor, maintaining the virus in a dormant state. Drug withdrawal restored reverse transcription, promoting efficient virus replication akin to that observed in latently infected patients on antiretroviral therapy. However, when HIV-1 MLOs were deliberately disassembled by pharmacological treatment, we observed a complete loss of viral infectivity. Our findings show that HIV-1 MLOs shield the final reverse transcription product from host immune detection. In vitro infection studies have shown that HIV-1 entry into the nucleus triggers the formation of dynamic structures called HIV-1 membraneless organelles (HIV-1-MLOs). This study reveals their existence in vivo and highlights their multifaceted role in the post-nuclear viral entry steps.Viral nuclear condensates form in a humanized mouse model after HIV-1 infection.Viral nuclear condensates host distinct viral core structures for several days post-infection.Nuclear condensates are the main sites of nuclear reverse transcription of the HIV-1 genome.Viral nuclear condensates protect the newly synthesized viral DNA from recognition by cGAS. HIV-1 membraneless organelles are the main sites of viral RNA reverse transcription in the host cell nucleus.