Quantitative Phosphoproteomics Reveals Extensive Cellular Reprogramming during HIV-1 Entry

• A quantitative phosphoproteomics screen of HIV-1-infected primary human CD4+ T cells
• The abundance of 239 phosphorylation sites from 175 genes changed after HIV-1 exposure
• Previously uncharacterized HIV-1 host factors, including SR proteins, confirmed by RNAi
• SRm300 and other SR proteins regulate alternative splicing of HIV-1 transcripts

SummaryReceptor engagement by HIV-1 during host cell entry activates signaling pathways that can reprogram the cell for optimal viral replication. To obtain a global view of the signaling events induced during HIV-1 entry, we conducted a quantitative phosphoproteomics screen of primary human CD4+ T cells after infection with an HIV-1 strain that engages the receptors CD4 and CXCR4. We quantified 1,757 phosphorylation sites with high stringency. The abundance of 239 phosphorylation sites from 175 genes, including several proteins in pathways known to be impacted by HIV-receptor binding, changed significantly within a minute after HIV-1 exposure. Several previously uncharacterized HIV-1 host factors were also identified and confirmed through RNAi depletion studies. Surprisingly, five serine/arginine-rich (SR) proteins involved in messenger RNA splicing, including the splicing factor SRm300 (SRRM2), were differentially phosophorylated. Mechanistic studies with SRRM2 suggest that HIV-1 modulates host cell alternative splicing machinery during entry in order to facilitate virus replication and release.