The vaccinia virus I1 protein is essential for the assembly of mature virions.
The product of the vaccinia virus I1 gene was characterized biochemically and genetically. This 35-kDa protein is conserved in diverse members of the poxvirus family but shows no homology to nonviral proteins. We show that recombinant I1 binds to both single-stranded and double-stranded DNA in a sequence-nonspecific manner in an electrophoretic mobility shift assay. The protein is expressed at late times during infection, and approximately 700 copies are encapsidated within the virion core. To determine the role of the I1 protein during the viral life cycle, a inducible viral recombinant in which the I1 gene was placed under the regulation of the Escherichia coli lac operator/repressor was constructed. In the absence of isopropyl-beta-D-thiogalactopyranoside, plaque formation was abolished and yields of infectious, intracellular virus were dramatically reduced. Although all phases of gene expression and DNA replication proceeded normally during nonpermissive infections, no mature virions were produced. Electron microscopic analysis confirmed the absence of mature virion assembly but revealed that apparently normal immature virions accumulated. Thus, I1 is an encapsidated DNA-binding protein required for the latest stages of vaccinia virion morphogenesis.
Klemperer
N
N
Department of Cell Biology and Anatomy, Cornell University Medical College, New York, New York 10021, USA.
Ward
J
J
Evans
E
E
Traktman
P
P
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D001483:Base Sequence; D002460:Cell Line; D004247:DNA; D004279:DNA, Viral; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D051379:Mice; D008969:Molecular Sequence Data; D010641:Phenotype; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D014616:Vaccinia virus / Q000235:genetics / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism; D014771:Virion / Q000502:physiology / Q000648:ultrastructure; D019065:Virus Assembly
D000914:Antibodies, Viral; D004279:DNA, Viral; D004268:DNA-Binding Proteins; C109350:I1 protein, vaccinia virus; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D014764:Viral Proteins; D004247:DNA; D010450:Endopeptidases