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Protein-RNA linkage and post-translational modifications of two sobemovirus VPgs

Allan Olspert1, Lauri Peil, Eugénie Hébrard

  • 1Department of Gene Technology, Tallinn University of Technology, Akadeemia tee 15, 12618 Tallinn, Estonia.

The Journal of General Virology
|November 12, 2010
PubMed

Insights

This study identifies the viral genome-linked protein (VPg) in Cocksfoot mottle virus (CfMV) and Rice yellow mottle virus (RYMV). Researchers found surprising variations in RNA-binding sites and phosphorylation patterns between these sobemoviruses.

Area of Science:

  • Virology
  • Molecular Biology
  • Protein Chemistry

Background:

  • Sobemoviruses feature a viral genome-linked protein (VPg) at the 5' end of their RNA genome.
  • VPg is generated through the processing of a larger viral polyprotein precursor.

Purpose of the Study:

  • To characterize the viral genome-linked proteins (VPgs) of Cocksfoot mottle virus (CfMV) and Rice yellow mottle virus (RYMV).
  • To determine the precise cleavage sites, termini, and post-translational modifications of CfMV and RYMV VPgs.

Main Methods:

  • Purification of VPgs from CfMV and RYMV virions.
  • Mass spectrometry analysis to identify cleavage sites, termini, and phosphorylation sites.
  • Experimental determination of VPg termini and covalent linkage to RNA.

Main Results:

  • Mature CfMV and RYMV VPgs were determined to be 78 and 79 amino acid residues long, respectively.
  • The amino acid residues covalently linked to RNA were not conserved, with Tyrosine at position 5 in CfMV VPg and Serine at position 1 in RYMV VPg.
  • Phosphorylation sites were identified in both VPgs, with similar locations (T20/S14 and S71/S72) and an additional site in RYMV VPg (S41).

Conclusions:

  • The study experimentally validated the termini and lengths of CfMV and RYMV VPgs.
  • Significant divergence in RNA-binding residues and distinct phosphorylation patterns were observed between CfMV and RYMV VPgs.
  • These findings provide crucial insights into the structural and functional diversity of VPgs within the Sobemovirus genus.

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