title: HIV-1 triggers WAVE2 phosphorylation in primary CD4 T cells and macrophages, mediating Arp2/3-dependent nuclear migration.
abstract: The human immunodeficiency virus type 1 (HIV-1) initiates receptor signaling and early actin dynamics during viral entry. This process is required for viral infection of primary targets such as resting CD4 T cells. WAVE2 is a component of a multiprotein complex linking receptor signaling to dynamic remodeling of the actin cytoskeleton. WAVE2 directly activates Arp2/3, leading to actin nucleation and filament branching. Although several bacterial and viral pathogens target Arp2/3 for intracellular mobility, it remains unknown whether HIV-1 actively modulates the Arp2/3 complex through virus-mediated receptor signal transduction. Here we report that HIV-1 triggers WAVE2 phosphorylation at serine 351 through gp120 binding to the chemokine coreceptor CXCR4 or CCR5 during entry. This phosphorylation event involves both Gαi-dependent and -independent pathways, and is conserved both in X4 and R5 viral infection of resting CD4 T cells and primary macrophages. We further demonstrate that inhibition of WAVE2-mediated Arp2/3 activity through stable shRNA knockdown of Arp3 dramatically diminished HIV-1 infection of CD4 T cells, preventing viral nuclear migration. Inhibition of Arp2/3 through a specific inhibitor, CK548, also drastically inhibited HIV-1 nuclear migration and infection of CD4 T cells. Our results suggest that Arp2/3 and the upstream regulator, WAVE2, are essential co-factors hijacked by HIV for intracellular migration, and may serve as novel targets to prevent HIV transmission.
authors.affiliation: National Center for Biodefense and Infectious Diseases, Department of Molecular and Microbiology, George Mason University, Manassas, Virginia 20110.
authors.forename: Mark
authors.initials: M
authors.lastname: Spear
authors.affiliation: National Center for Biodefense and Infectious Diseases, Department of Molecular and Microbiology, George Mason University, Manassas, Virginia 20110.
authors.forename: Jia
authors.initials: J
authors.lastname: Guo
authors.affiliation: National Center for Biodefense and Infectious Diseases, Department of Molecular and Microbiology, George Mason University, Manassas, Virginia 20110.
authors.forename: Amy
authors.initials: A
authors.lastname: Turner
authors.affiliation: National Center for Biodefense and Infectious Diseases, Department of Molecular and Microbiology, George Mason University, Manassas, Virginia 20110.
authors.forename: Dongyang
authors.initials: D
authors.lastname: Yu
authors.affiliation: National Center for Biodefense and Infectious Diseases, Department of Molecular and Microbiology, George Mason University, Manassas, Virginia 20110.
authors.forename: Weifeng
authors.initials: W
authors.lastname: Wang
authors.affiliation: National Center for Biodefense and Infectious Diseases, Department of Molecular and Microbiology, George Mason University, Manassas, Virginia 20110.
authors.forename: Beatrix
authors.initials: B
authors.lastname: Meltzer
authors.affiliation: Key Laboratory of Immunology of AIDS, Ministry of Health, the First Affiliated Hospital, China Medical University, Shenyang, Liaoning province 110001, China.
authors.forename: Sijia
authors.initials: S
authors.lastname: He
authors.affiliation: Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia 24060.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Hu
authors.affiliation: Key Laboratory of Immunology of AIDS, Ministry of Health, the First Affiliated Hospital, China Medical University, Shenyang, Liaoning province 110001, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Shang
authors.affiliation: Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia 24060.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Kuhn
authors.affiliation: National Center for Biodefense and Infectious Diseases, Department of Molecular and Microbiology, George Mason University, Manassas, Virginia 20110. Electronic address: ywu8@gmu.edu.
authors.forename: Yuntao
authors.initials: Y
authors.lastname: Wu
mesh_terms: D051376:Actin-Related Protein 2-3 Complex / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism / Q000821:virology*; D002467:Cell Nucleus / Q000821:virology*; D055785:Gene Knockdown Techniques; D015658:HIV Infections / Q000235:genetics / Q000378:metabolism* / Q000821:virology*; D015497:HIV-1 / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000821:virology*; D010766:Phosphorylation; D034741:RNA, Small Interfering / Q000235:genetics; D014779:Virus Replication; D051300:Wiskott-Aldrich Syndrome Protein Family / Q000378:metabolism*
chemical_list: D051376:Actin-Related Protein 2-3 Complex; D034741:RNA, Small Interfering; C120152:WASF2 protein, human; D051300:Wiskott-Aldrich Syndrome Protein Family
keywords: Arp2/3; CCR5; CD4; HIV-1; Immunology; Microbiology; Signal Transduction; Virology; Wav2