The inability of vaccinia virus A33R protein to form intermolecular disulfide-bonded homodimers does not affect the production of infectious extracellular virus.
The orthopoxvirus protein A33 forms a disulfide-bonded high molecular weight species that could be either a homodimer or a heteromultimer. The protein is a major target for neutralizing antibodies and the majority of antibodies raised against A33 only recognize the disulfide-bonded form. Here, we report that A33 is present as a disulfide-bonded homodimer during infection. Additionally, we examined the function of intermolecular disulfide bonding in A33 homodimerization during infection. We show that the cysteine at amino acid 62 is required for intermolecular disulfide bonding, but not dimerization as this mutant was still able to form homodimers. To investigate the role of disulfide-bonded homodimers during viral morphogenesis, recombinant viruses that express an A33R with cysteine 62 mutated to serine were generated. The recombinant viruses had growth characteristics similar to their parental viruses, indicating that intermolecular disulfide-bonded homodimerization of A33 is not required for its function.
Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY 14642, USA.
Winnie M
WM
Chan
Aja E
AE
Kalkanoglu
Brian M
BM
Ward
D003545:Cysteine / Q000235:genetics / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D055503:Protein Multimerization; D012694:Serine / Q000235:genetics / Q000378:metabolism; D014616:Vaccinia virus / Q000254:growth & development / Q000472:pathogenicity / Q000502:physiology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D019562:Viral Load; D010948:Viral Plaque Assay; D019065:Virus Assembly; D014779:Virus Replication
C446940:A33R protein, Vaccinia virus; D004220:Disulfides; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D012694:Serine; D003545:Cysteine