Characterization of Simian Immunodeficiency Virus SIVSM/Human Immunodeficiency Virus Type 2 Vpx Function in Human Myeloid Cells

ABSTRACT Human immunodeficiency virus type 2 (HIV-2)/simian immunodeficiency virus SIVSM Vpx is incorporated into virion particles and is thus present during the early steps of infection, when it has been reported to influence the nuclear import of viral DNA. We recently reported that Vpx promoted the accumulation of full-length viral DNA following the infection of human monocyte-derived dendritic cells (DCs). This positive effect was exerted following the infection of DCs with cognate viruses and with retroviruses as divergent as HIV-1, feline immunodeficiency virus, and even murine leukemia virus, leading us to suggest that Vpx counteracted an antiviral restriction present in DCs. Here, we show that Vpx is required, albeit to a different extent, for the infection of all myeloid but not of lymphoid cells, including monocytes, macrophages, and monocytoid THP-1 cells that had been induced to differentiate with phorbol esters. The intracellular localization of Vpx was highly heterogeneous and cell type dependent, since Vpx localized differently in HeLa cells and DCs. Despite these differences, no clear correlation between the functionality of Vpx and its intracellular localization could be drawn. As a first insight into its function, we determined that SIVSM/HIV-2 and SIVRCM Vpx proteins interact with the DCAF1 adaptor of the Cul4-based E3 ubiquitin ligase complex recently described to associate with HIV-1 Vpr and HIV-2 Vpx. However, the functionality of Vpx proteins in the infection of DCs did not strictly correlate with DCAF1 binding, and knockdown experiments failed to reveal a functional role for this association in differentiated THP-1 cells. Lastly, when transferred in the context of a replication-competent viral clone, Vpx was required for replication in DCs.

[1]  Y. Kotake,et al.  Human Immunodeficiency Virus Type 1 Vpr-Binding Protein VprBP, a WD40 Protein Associated with the DDB1-CUL4 E3 Ubiquitin Ligase, Is Essential for DNA Replication and Embryonic Development , 2008, Molecular and Cellular Biology.

[2]  M. Otsuka,et al.  Vpx Is Critical for Reverse Transcription of the Human Immunodeficiency Virus Type 2 Genome in Macrophages , 2008, Journal of Virology.

[3]  Smita Srivastava,et al.  Lentiviral Vpx Accessory Factor Targets VprBP/DCAF1 Substrate Adaptor for Cullin 4 E3 Ubiquitin Ligase to Enable Macrophage Infection , 2008, PLoS pathogens.

[4]  Rajnish Kaushik,et al.  Primate Lentiviral Vpx Commandeers DDB1 to Counteract a Macrophage Restriction , 2008, PLoS pathogens.

[5]  J. Darlix,et al.  Characterization of the Early Steps of Infection of Primary Blood Monocytes by Human Immunodeficiency Virus Type 1 , 2008, Journal of Virology.

[6]  L. Ratner,et al.  Hsp40 Facilitates Nuclear Import of the Human Immunodeficiency Virus Type 2 Vpx-Mediated Preintegration Complex , 2007, Journal of Virology.

[7]  C. D. de Noronha,et al.  The HIV1 Protein Vpr Acts to Promote G2 Cell Cycle Arrest by Engaging a DDB1 and Cullin4A-containing Ubiquitin Ligase Complex Using VprBP/DCAF1 as an Adaptor* , 2007, Journal of Biological Chemistry.

[8]  K. Cichutek,et al.  Impact of viral accessory proteins of SIVsmmPBj on early steps of infection of quiescent cells. , 2007, Virology.

[9]  Xiao-Fang Yu,et al.  DDB1 and Cul4A Are Required for Human Immunodeficiency Virus Type 1 Vpr-Induced G2 Arrest , 2007, Journal of Virology.

[10]  M. Washburn,et al.  Lentiviral Vpr usurps Cul4–DDB1[VprBP] E3 ubiquitin ligase to modulate cell cycle , 2007, Proceedings of the National Academy of Sciences.

[11]  A. Finzi,et al.  HIV-1 Vpr-Mediated G2 Arrest Involves the DDB1-CUL4AVPRBP E3 Ubiquitin Ligase , 2007, PLoS pathogens.

[12]  E. Zimmerman,et al.  HIV-1 Vpr activates the G2 checkpoint through manipulation of the ubiquitin proteasome system , 2007, Virology Journal.

[13]  N. Landau,et al.  HIV-1 Vpr function is mediated by interaction with the damage-specific DNA-binding protein DDB1 , 2007, Proceedings of the National Academy of Sciences.

[14]  Jean-Christophe Rain,et al.  HIV1 Vpr Arrests the Cell Cycle by Recruiting DCAF1/VprBP, a Receptor of the Cul4-DDB1 Ubiquitin Ligase , 2007, Cell cycle.

[15]  T. Hashikawa,et al.  Cell-cycle-dependent dynamics of nuclear pores: pore-free islands and lamins , 2006, Journal of Cell Science.

[16]  S. Mahalingam,et al.  Nuclear Export of Simian Immunodeficiency Virus Vpx Protein , 2006, Journal of Virology.

[17]  J. Darlix,et al.  With a little help from a friend: increasing HIV transduction of monocyte-derived dendritic cells with virion-like particles of SIVMAC , 2006, Gene Therapy.

[18]  L. Ratner,et al.  Analysis of HIV-2 Vpx by modeling and insertional mutagenesis. , 2006, Virology.

[19]  L. Ratner,et al.  Conserved amino acids of the human immunodeficiency virus type 2 Vpx nuclear localization signal are critical for nuclear targeting of the viral preintegration complex in non-dividing cells. , 2006, Virology.

[20]  S. Mahalingam,et al.  Phosphorylation by MAPK Regulates Simian Immunodeficiency Virus Vpx Protein Nuclear Import and Virus Infectivity* , 2005, Journal of Biological Chemistry.

[21]  J. Darlix,et al.  Heterologous Human Immunodeficiency Virus Type 1 Lentiviral Vectors Packaging a Simian Immunodeficiency Virus-Derived Genome Display a Specific Postentry Transduction Defect in Dendritic Cells , 2003, Journal of Virology.

[22]  L. Ratner,et al.  Identification of the nuclear localization signal of human immunodeficiency virus type 2 Vpx. , 2003, Virology.

[23]  H. Shiota,et al.  Vpx and Vpr proteins of HIV-2 up-regulate the viral infectivity by a distinct mechanism in lymphocytic cells. , 2003, Microbes and infection.

[24]  B. Roques,et al.  NMR structure of the HIV-1 regulatory protein VPR. , 2003, Journal of molecular biology.

[25]  P. Mangeot,et al.  High levels of transduction of human dendritic cells with optimized SIV vectors. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[26]  Jan Ellenberg,et al.  Nuclear pore complexes form immobile networks and have a very low turnover in live mammalian cells , 2001, The Journal of cell biology.

[27]  G. Shaw,et al.  Functional Analysis of the Simian Immunodeficiency Virus Vpx Protein: Identification of Packaging Determinants and a Novel Nuclear Targeting Domain , 2001, Journal of Virology.

[28]  C. Depienne,et al.  Cellular distribution and karyophilic properties of matrix, integrase, and Vpr proteins from the human and simian immunodeficiency viruses. , 2000, Experimental cell research.

[29]  L. Ratner,et al.  The C-Terminal Proline-Rich Tail of Human Immunodeficiency Virus Type 2 Vpx Is Necessary for Nuclear Localization of the Viral Preintegration Complex in Nondividing Cells , 2000, Journal of Virology.

[30]  Morris S. Jones,et al.  A Conserved Dileucine-Containing Motif in p6gag Governs the Particle Association of Vpx and Vpr of Simian Immunodeficiency Viruses SIVmac and SIVagm , 1999, Journal of Virology.

[31]  L. Selig,et al.  Interaction with the p6 Domain of the Gag Precursor Mediates Incorporation into Virions of Vpr and Vpx Proteins from Primate Lentiviruses , 1999, Journal of Virology.

[32]  J. Wakefield,et al.  Vpx is required for dissemination and pathogenesis of SIVSM PBj: Evidence of macrophage-dependent viral amplification , 1998, Nature Medicine.

[33]  P. Sharp,et al.  Nuclear import and cell cycle arrest functions of the HIV‐1 Vpr protein are encoded by two separate genes in HIV‐2/SIV(SM). , 1996, The EMBO journal.

[34]  V. Cordes,et al.  Cytoplasmic annulate lamellae in cultured cells: composition, distribution, and mitotic behavior , 1996, Cell and Tissue Research.

[35]  F. Gage,et al.  In Vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector , 1996, Science.

[36]  J. Sodroski,et al.  Functional analysis of the vpx, vpr, and nef genes of simian immunodeficiency virus. , 1995, Journal of acquired immune deficiency syndromes and human retrovirology : official publication of the International Retrovirology Association.

[37]  A. Adachi,et al.  Human Immunodeficiency Virus Vpx Is Required for the Early Phase of Replication in Peripheral Blood Mononuclear Cells , 1994, Microbiology and immunology.

[38]  J. Kappes,et al.  Localization of the Vpx packaging signal within the C terminus of the human immunodeficiency virus type 2 Gag precursor protein , 1994, Journal of virology.

[39]  R. Desrosiers,et al.  Construction and in vitro properties of SIVmac mutants with deletions in "nonessential" genes. , 1994, AIDS research and human retroviruses.

[40]  J. Kappes,et al.  Intracellular transport and virion incorporation of vpx requires interaction with other virus type-specific components. , 1993, Virology.

[41]  M. Tristem,et al.  Evolution of the primate lentiviruses: evidence from vpx and vpr. , 1992, The EMBO journal.

[42]  J. Kappes,et al.  Human immunodeficiency virus type 2 vpx protein augments viral infectivity. , 1991, Virology.

[43]  X. Yu,et al.  The vpx gene of simian immunodeficiency virus facilitates efficient viral replication in fresh lymphocytes and macrophage , 1991, Journal of virology.

[44]  M. Emerman,et al.  VPX mutants of HIV‐2 are infectious in established cell lines but display a severe defect in peripheral blood lymphocytes. , 1989, The EMBO journal.

[45]  J. Darlix,et al.  SIVSM / HIV-2 Vpx proteins promote retroviral escape from a proteasome-dependent restriction pathway present in human dendritic cells , 2007 .

[46]  Y. Takebe,et al.  Biological characterization of human immunodeficiency virus type 1 and type 2 mutants in human peripheral blood mononuclear cells , 2005, Archives of Virology.