title: Functional analysis of Vif protein shows less restriction of human immunodeficiency virus type 2 by APOBEC3G.
abstract: Viral infectivity factor (Vif) is one of the human immunodeficiency virus (HIV) accessory proteins and is conserved in the primate lentivirus group. This protein is essential for viral replication in vivo and for productive infection of nonpermissive cells, such as peripheral blood mononuclear cells (PBMC). Vif counteracts an antiretroviral cellular factor in nonpermissive cells named CEM15/APOBEC3G. Although HIV type 1 (HIV-1) Vif protein (Vif1) can be functionally replaced by HIV-2 Vif protein (Vif2), its identity is very small. Most of the functional studies have been carried out with Vif1. Characterization of functional domains of Vif2 may elucidate its function, as well as differences between HIV-1 and HIV-2 infectivity. Our aim was to identify the permissivity of different cell lines for HIV-2 vif-minus viruses. By mutagenesis specific conserved motifs of HIV-2 Vif protein were analyzed, as well as in conserved motifs between Vif1 and Vif2 proteins. Vif2 mutants were examined for their stability, expression, and cellular localization in order to characterize essential domains of Vif2 proteins. Viral replication in various target cells (PBMC and H9, A3.01, U38, and Jurkat cells) and infectivity in single cycle assays in the presence of APOBEC3G were also analyzed. Our results of viral replication show that only PBMC have a nonpermissive phenotype in the absence of Vif2. Moreover, the HIV-1 vif-minus nonpermissive cell line H9 does not show a similar phenotype for vif-negative HIV-2. We also report a limited effect of APOBEC3G in a single-cycle infectivity assay, where only conserved domains between HIV-1 and HIV-2 Vif proteins influence viral infectivity. Taken together, these results allow us to speculate that viral inhibition by APOBEC3G is not the sole and most important determinant of antiviral activity against HIV-2.
authors.affiliation: Instituto Superior de Ciências da Saúde-Sul, Quinta da Granja, Monte da Caparica, 2829-511 Caparica, Portugal.
authors.forename: Ana Clara
authors.initials: AC
authors.lastname: Ribeiro
authors.forename: Alexandra
authors.initials: A
authors.lastname: Maia e Silva
authors.forename: Mariana
authors.initials: M
authors.lastname: Santa-Marta
authors.forename: Ana
authors.initials: A
authors.lastname: Pombo
authors.forename: José
authors.initials: J
authors.lastname: Moniz-Pereira
authors.forename: Joao
authors.initials: J
authors.lastname: Goncalves
authors.forename: Isabel
authors.initials: I
authors.lastname: Barahona
mesh_terms: D000071480:APOBEC-3G Deaminase; D000595:Amino Acid Sequence; D003564:Cytidine Deaminase; D016346:Gene Products, vif / Q000737:chemistry / Q000502:physiology*; D015498:HIV-2 / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009700:Nucleoside Deaminases; D011506:Proteins / Q000502:physiology*; D012097:Repressor Proteins; D014779:Virus Replication; D054320:vif Gene Products, Human Immunodeficiency Virus
chemical_list: D016346:Gene Products, vif; D011506:Proteins; D012097:Repressor Proteins; D054320:vif Gene Products, Human Immunodeficiency Virus; D009700:Nucleoside Deaminases; D000071480:APOBEC-3G Deaminase; C464924:APOBEC3G protein, human; D003564:Cytidine Deaminase