title: Molecular insight into how HIV-1 Vpr protein impairs cell growth through two genetically distinct pathways.
abstract: Vpr, a small HIV auxiliary protein, hijacks the CUL4 ubiquitin ligase through DCAF1 to inactivate an unknown cellular target, leading to cell cycle arrest at the G(2) phase and cell death. Here we first sought to delineate the Vpr determinants involved in the binding to DCAF1 and to the target. On the one hand, the three α-helices of Vpr are necessary and sufficient for binding to DCAF1; on the other hand, nonlinear determinants in Vpr are required for binding to the target, as shown by using protein chimeras. We also underscore that a SRIG motif conserved in the C-terminal tail of Vpr proteins from HIV-1/SIVcpz and HIV-2/SIVsmm lineages is critical for G(2) arrest. Our results suggest that this motif may be predictive of the ability of Vpr proteins from other SIV lineages to mediate G(2) arrest. We took advantage of the characterization of a subset of G(2) arrest-defective, but DCAF1 binding-proficient mutants, to investigate whether Vpr interferes with cell viability independently of its ability to induce G(2) arrest. These mutants inhibited cell colony formation in HeLa cells and are cytotoxic in lymphocytes, unmasking a G(2) arrest-independent cytopathic effect of Vpr. Furthermore these mutants do not block cell cycle progression at the G(1) or S phases but trigger apoptosis through caspase 3. Disruption of DCAF1 binding restored efficiency of colony formation. However, DCAF1 binding per se is not sufficient to confer cytopathicity. These data support a model in which Vpr recruits DCAF1 to induce the degradation of two host proteins independently required for proper cell growth.
authors.affiliation: Inserm, U1016, Institut Cochin, Paris 75014, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Maudet
authors.forename: Matthieu
authors.initials: M
authors.lastname: Bertrand
authors.forename: Erwann
authors.initials: E
authors.lastname: Le Rouzic
authors.forename: Hichem
authors.initials: H
authors.lastname: Lahouassa
authors.forename: Diana
authors.initials: D
authors.lastname: Ayinde
authors.forename: Sébastien
authors.initials: S
authors.lastname: Nisole
authors.forename: Caroline
authors.initials: C
authors.lastname: Goujon
authors.forename: Andrea
authors.initials: A
authors.lastname: Cimarelli
authors.forename: Florence
authors.initials: F
authors.lastname: Margottin-Goguet
authors.forename: Catherine
authors.initials: C
authors.lastname: Transy
mesh_terms: D020816:Amino Acid Motifs; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002453:Cell Cycle; D016923:Cell Death / Q000235:genetics; D044842:Cullin Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D009154:Mutation; D017346:Protein Serine-Threonine Kinases; D044767:Ubiquitin-Protein Ligases; D054325:vpr Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D044842:Cullin Proteins; D054325:vpr Gene Products, Human Immunodeficiency Virus; D044767:Ubiquitin-Protein Ligases; C530533:DCAF1 protein, human; D017346:Protein Serine-Threonine Kinases