The extracellular domain of vaccinia virus protein B5R affects plaque phenotype, extracellular enveloped virus release, and intracellular actin tail formation.
Vaccinia virus produces two morphologically distinct forms of infectious virus, termed intracellular mature virus (IMV) and extracellular enveloped virus (EEV). EEV is important for virus dissemination within a host and has different surface proteins which bind to cell receptors different from those used by IMV. Six genes are known to encode EEV-specific proteins. One of these, B5R, encodes a 42-kDa glycoprotein with amino acid similarity to members of the complement control protein superfamily and contains four copies of a 50- to 70-amino-acid repeat called the short consensus repeat (SCR). Deletion of B5R causes a small-plaque phenotype, a 10-fold reduction in EEV formation, and virus attenuation in vivo. In this study, we inserted mutated versions of the B5R gene lacking different combinations of the SCRs into a virus deletion mutant lacking the B5R gene. The resultant viruses each formed small plaques only slightly larger than those of the deletion mutant; however, the virus containing only SCR 1 formed plaques slightly larger than those of viruses with SCRs 1 and 2 or SCRs 1, 2, and 3. All of these viruses produced approximately 50-fold more infectious EEV than wild-type virus and formed comet-shaped plaques under liquid overlay. Despite producing more EEV, the mutant viruses were unable to induce the polymerization of actin on intracellular virus particles. The implications of these results for our understanding of EEV formation, release, and infectivity are discussed.
Mathew
E
E
Sir William Dunn School of Pathology, University of Oxford, United Kingdom.
Sanderson
C M
CM
Hollinshead
M
M
Smith
G L
GL
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002522:Chlorocebus aethiops; D016384:Consensus Sequence; D006801:Humans; D007425:Intracellular Membranes; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D016296:Mutagenesis; D010641:Phenotype; D011817:Rabbits; D012091:Repetitive Sequences, Nucleic Acid; D017384:Sequence Deletion; D014616:Vaccinia virus / Q000235:genetics / Q000254:growth & development / Q000502:physiology* / Q000648:ultrastructure; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000032:analysis; D019065:Virus Assembly
D000199:Actins; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C074512:42kDa protein, Vaccinia virus