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Structural characterization of HC-Pro, a plant virus multifunctional protein

Authors
  • Plisson, Célia
  • Drucker, Martin
  • Blanc, Stéphane
  • German-Retana, Sylvie
  • Le Gall, Olivier
  • Thomas, Daniel
  • Bron, Patrick
Publication Date
Jan 01, 2003
Source
ProdInra
Keywords
Language
English
License
Unknown
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Abstract

The helper component proteinase (HC-Pro) is a key protein encoded by plant viruses of the genus Potyvirus. HC-Pro is involved in different steps of the viral cycle, aphid transmission, replication, and virus cell-to-cell and systemic movement and is a suppressor of post-transcriptional gene silencing. Structural knowledge of HC-Pro is required to better understand its multiple functions. To this aim, we purified His-tagged wild-type HC-Pro and a N-terminal deletion mutant (ΔHC-Pro) from plants infected with recombinant potyviruses. Biochemical analysis of the recombinant proteins confirmed that HC-Pro is a dimer in solution, that the N terminus is not essential for self-interaction, and that a large C-terminal domain is highly resistant to proteolysis. Two-dimensional crystals of the recombinant proteins were successfully grown on Ni2+-chelating lipid monolayers. Comparison of projection maps of negatively stained crystals revealed that HC-Pro is composed of two domains separated by a flexible constriction. Cryo-electron crystallography of ΔHC-Pro allowed us to calculate a projection map at 9-Å resolution. Our data from electron microscopy, biochemical analysis, and secondary structure predictions lead us to suggest a model for structure/function relationships in the HC-Pro protein.

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