title: Quantitative phosphoproteomics reveals extensive cellular reprogramming during HIV-1 entry.
abstract: Receptor 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.
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia Research Institute, Philadelphia, PA 19104, USA.
authors.forename: Jason A
authors.initials: JA
authors.lastname: Wojcechowskyj
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia Research Institute, Philadelphia, PA 19104, USA.
authors.forename: Chuka A
authors.initials: CA
authors.lastname: Didigu
authors.affiliation: Protein and Proteomics Core, Children's Hospital of Philadelphia Research Institute, Philadelphia, PA 19104, USA.
authors.forename: Jessica Y
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Nicholas F
authors.initials: NF
authors.lastname: Parrish
authors.affiliation: Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Rohini
authors.initials: R
authors.lastname: Sinha
authors.affiliation: Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Beatrice H
authors.initials: BH
authors.lastname: Hahn
authors.affiliation: Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Frederic D
authors.initials: FD
authors.lastname: Bushman
authors.affiliation: Department of Statistics, The Wharton School, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Shane T
authors.initials: ST
authors.lastname: Jensen
authors.affiliation: Protein and Proteomics Core, Children's Hospital of Philadelphia Research Institute, Philadelphia, PA 19104, USA.
authors.forename: Steven H
authors.initials: SH
authors.lastname: Seeholzer
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia Research Institute, Philadelphia, PA 19104, USA. Electronic address: domsr@email.chop.edu.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Doms
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000821:virology*; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D015497:HIV-1 / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D010750:Phosphoproteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D012326:RNA Splicing; D053586:Virus Internalization; D057074:Virus Release; D014779:Virus Replication
chemical_list: D010750:Phosphoproteins; D020543:Proteome