Assembly of the Hv190S totivirus capsid is independent of posttranslational modification of the capsid protein.
The genome of Helminthosporium victoriae 190S totivirus (Hv190SV) consists of two large overlapping open reading frames (ORFs), encoding a capsid protein (CP) and an RNA-dependent RNA polymerase. The capsid of Hv190SV, even though encoded by a single gene, contains three closely related capsid polypeptides: p88, p83, and p78. p88 and p83 are phosphorylated, whereas p78, which is derived from p88 via proteolytic processing at the C terminus, is nonphosphorylated. In this study we expressed the CP ORF in bacteria and determined that a single product comigrating with virion p88 was generated. Evidence from in vivo phosphorylation studies indicated that the bacterially expressed p88 was unmodified, and thus autophosphorylation was ruled out. Enzymatic-dephosphorylation experiments using 32P-labeled p88 as a substrate demonstrated that the phosphorylated and nonphosphorylated forms of p88 could not be differentiated based on their mobilities in SDS gels and suggested that the two forms occur in purified virions. We also showed that the unmodified p88 is competent for assembly into virus-like particles, indicating that neither phosphorylation nor proteolytic processing of CP is required for capsid assembly. Posttranslational modification of CP, however, is proposed to play an important role in the life cycle of Hv190SV, including regulation of transcription/replication and/or packaging/release from virions of the viral (+) strand RNA transcript.
Soldevila
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AI
Department of Plant Pathology, University of Kentucky, Lexington, Kentucky, 40546-0091, USA.
Huang
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S
Ghabrial1
S A
SA
D002213:Capsid / Q000378:metabolism*; D036022:Capsid Proteins; D004279:DNA, Viral / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000648:ultrastructure / Q000821:virology; D015870:Gene Expression; D006375:Helminthosporium / Q000821:virology*; D008854:Microscopy, Electron; D016366:Open Reading Frames; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D019197:Totivirus / Q000235:genetics*; D014771:Virion / Q000378:metabolism; D019065:Virus Assembly
D036022:Capsid Proteins; D004279:DNA, Viral; C102211:capsid protein p88, Hv190SV