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The hypervariable amino-terminus of P1 protease modulates potyviral replication and host defense responses

PLoS Pathog. 2014 Mar 6;10(3):e1003985. doi: 10.1371/journal.ppat.1003985. eCollection 2014 Mar.

Abstract

The replication of many RNA viruses involves the translation of polyproteins, whose processing by endopeptidases is a critical step for the release of functional subunits. P1 is the first protease encoded in plant potyvirus genomes; once activated by an as-yet-unknown host factor, it acts in cis on its own C-terminal end, hydrolyzing the P1-HCPro junction. Earlier research suggests that P1 cooperates with HCPro to inhibit host RNA silencing defenses. Using Plum pox virus as a model, we show that although P1 does not have a major direct role in RNA silencing suppression, it can indeed modulate HCPro function by its self-cleavage activity. To study P1 protease regulation, we used bioinformatic analysis and in vitro activity experiments to map the core C-terminal catalytic domain. We present evidence that the hypervariable region that precedes the protease domain is predicted as intrinsically disordered, and that it behaves as a negative regulator of P1 proteolytic activity in in vitro cleavage assays. In viral infections, removal of the P1 protease antagonistic regulator is associated with greater symptom severity, induction of salicylate-dependent pathogenesis-related proteins, and reduced viral loads. We suggest that fine modulation of a viral protease activity has evolved to keep viral amplification below host-detrimental levels, and thus to maintain higher long-term replicative capacity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Blotting, Western
  • Chromatography, Liquid
  • Host-Parasite Interactions / physiology*
  • Molecular Sequence Data
  • Nicotiana / virology
  • Peptide Hydrolases / chemistry
  • Peptide Hydrolases / metabolism*
  • Plant Diseases / virology
  • Potyvirus / pathogenicity
  • Potyvirus / physiology*
  • Protein Structure, Tertiary / physiology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Tandem Mass Spectrometry
  • Viral Proteins / chemistry
  • Viral Proteins / metabolism*
  • Virus Replication / physiology*

Substances

  • Viral Proteins
  • Peptide Hydrolases

Grants and funding

This work was funded by grants BIO2010-18541 from the Spanish “Ministerio de Ciencia e Innovación”, and KBBE-204429 from the European Union. FP is financed by a La Caixa PhD fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.