Mutational analysis of the feline immunodeficiency virus matrix protein.
To study the process of feline immunodeficiency virus (FIV) assembly, we examined the suitability of the vaccinia vector system to reproduce FIV particle formation. To this end, we constructed a recombinant vaccinia virus carrying the FIV gag gene. Biochemical and electron microscopy analyses of cells infected with this recombinant virus showed that the FIV Gag polyprotein self-assembled into lentivirus-like particles that were released into the culture medium. As a first step in the identification of molecular determinants in FIV Gag that are involved in virus assembly, we performed a site-directed mutagenesis analysis of the N-terminal matrix (MA) domain of the FIV Gag precursor. To this end, a series of amino acid substitutions and small in-frame deletions were introduced into the FIV MA and the mutated FIV gag gene constructs were expressed by means of the vaccinia system. Characterization of the assembly phenotype of these FIV Gag mutants led to the identification of amino acidic regions within the MA domain that are necessary for efficient transport of the Gag precursor to the plasma membrane and particle assembly. Our results reveal the role that the FIV MA plays in virus morphogenesis and contribute to the understanding of the assembly process in non-primate lentiviruses.
Manrique
M L
ML
Centro de Virología Animal (CEVAN-CONICET), Serrano 669, C1414DEM, Buenos Aires, Argentina.
Celma
C C
CC
González
S A
SA
Affranchino
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D004274:DNA, Recombinant / Q000235:genetics; D005347:Fibroblasts; D015683:Gene Products, gag / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015750:Genes, gag / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D016089:Immunodeficiency Virus, Feline / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D017434:Protein Structure, Tertiary; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019065:Virus Assembly
D004274:DNA, Recombinant; D015683:Gene Products, gag; D014763:Viral Matrix Proteins; D013937:Thymidine Kinase