Physical and functional interactions between vaccinia virus F10 protein kinase and virion assembly proteins A30 and G7.
An early step in vaccinia virus morphogenesis, the association of crescent membranes with electron-dense granular material, is perturbed when expression of the viral protein encoded by the A30L or G7L open reading frame is repressed. Under these conditions, we found that phosphorylation of the A17 membrane protein, which is mediated by the F10 kinase, was severely reduced. Furthermore, A30 and G7 stimulated F10-dependent phosphorylation of A17 in the absence of other viral late proteins. Evidence for physical interactions between A30, G7, and F10 was obtained by their coimmunoprecipitation with antibody against A30 or F10. In addition, phosphorylation of A30 was dependent on the F10 kinase and autophosphorylation of F10 was stimulated by A30 and G7. Nevertheless, the association of A30, G7, and F10 occurred even with mutated, catalytically inactive forms of F10. Just as A30 and G7 are mutually dependent on each other for stability, F10 was nearly undetectable in the absence of A30 and G7. The reverse is not true, however, as repression of F10 did not diminish A30 or G7. Interaction of F10 with A30 and G7 presumably occurred within the virus factory areas of the cytoplasm, where each was concentrated. F10 localized predominantly in the cortical region of immature virions, beneath the membrane where A17 is located. F10 remained associated with the particulate core fraction of mature virions after treatment with a nonionic detergent and reducing agent. The formation of protein complexes such as the one involving A30, G7, and F10 may be a mechanism for the regulated packaging and processing of virion components.
Szajner
Patricia
P
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
Weisberg
Andrea S
AS
Moss
Bernard
B
D000818:Animals; D002460:Cell Line; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D016253:Microscopy, Immunoelectron; D010766:Phosphorylation; D011233:Precipitin Tests; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism* / Q000502:physiology; D011817:Rabbits; D013912:Threonine / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism; D014616:Vaccinia virus / Q000235:genetics / Q000378:metabolism*; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D014764:Viral Proteins / Q000378:metabolism* / Q000502:physiology; D014771:Virion / Q000378:metabolism*; D019065:Virus Assembly
D014758:Viral Core Proteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D013912:Threonine; D014443:Tyrosine; C095546:protein kinase 2, Vaccinia virus; D017346:Protein Serine-Threonine Kinases