title: Mutational analysis of the conserved cysteine residues in the simian immunodeficiency virus matrix protein.
abstract: The matrix protein (MA) of human and simian immunodeficiency viruses (HIV and SIV) is encoded by the amino-terminal region of the Gag precursor and has been suggested to be involved in different processes during the early and late stages of the virus life cycle. The MA protein of SIV contains three cysteine residues at positions 57, 83, and 87, which are also highly conserved among HIV-2 isolates. In order to study the functional significance of these residues in virus morphogenesis, a series of mutations affecting the cysteines of SIV MA were introduced into a gag-protease construct and expressed in the vaccinia vector system. The MA mutants were assayed for their ability to synthesize and process the Gag polyprotein precursor as well as to release particles into the culture medium. In addition, the incorporation of the envelope glycoprotein (Env) into the Gag-made particles was investigated. Substitution of alanine for cysteine 87 had little effect on particle release and Env glycoprotein association. By contrast, the individual replacement of cysteines 57 or 83 by alanine, as well as the simultaneous mutation of cysteines 83 and 87, significantly reduced the ability of Gag polypeptides to produce extracellular particles. Assembly into particles appeared to be also affected, albeit to a lesser extent, when both cysteines 57 and 83 were replaced by alanine. Furthermore, substitution of cysteine 83 in the SIV MA domain was found to be detrimental to Gag polyprotein processing. Analysis of the Env glycoprotein association with recombinant particles revealed that this process was moderately affected in the case of the double mutants lacking cysteines 57 and 83, or cysteines 57 and 87, and the cysteine-minus triple mutant. Our results suggest that the conserved cysteines 57 and 83 in the MA domain are important for efficient SIV Gag particle production.
authors.affiliation: Centro de Virología Animal (CEVAN-CONICET), Buenos Aires, Argentina.
authors.forename: S A
authors.initials: SA
authors.lastname: González
authors.forename: J L
authors.initials: JL
authors.lastname: Affranchino
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D017124:Conserved Sequence / Q000235:genetics*; D003545:Cysteine / Q000502:physiology*; D004252:DNA Mutational Analysis; D015686:Gene Products, env / Q000378:metabolism; D015683:Gene Products, gag / Q000096:biosynthesis / Q000378:metabolism; D015750:Genes, gag / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D015699:HIV Envelope Protein gp120 / Q000032:analysis; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D009024:Morphogenesis; D009154:Mutation / Q000502:physiology; D011499:Protein Processing, Post-Translational; D015302:Simian Immunodeficiency Virus / Q000235:genetics / Q000502:physiology*; D014616:Vaccinia virus / Q000235:genetics; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins / Q000235:genetics* / Q000502:physiology; D014771:Virion; D014779:Virus Replication / Q000235:genetics*
chemical_list: D015686:Gene Products, env; D015683:Gene Products, gag; D015699:HIV Envelope Protein gp120; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; C069623:gp120 protein, Simian immunodeficiency virus; D003545:Cysteine