Sterol binding by the tombusviral replication proteins is essential for replication in yeast and plants

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31 Scopus citations

Abstract

Membranous structures derived from various organelles are important for replication of plus-stranded RNA viruses. Although the important roles of coopted host proteins in RNA virus replication have been appreciated for a decade, the equally important functions of cellular lipids in virus replication have been gaining full attention only recently. Previous work with Tomato bushy stunt tombusvirus (TBSV) in model host yeast has revealed essential roles for phosphatidylethanolamine and sterols in viral replication. To further our understanding of the role of sterols in tombusvirus replication, in this work we showed that the TBSV p33 and p92 replication proteins could bind to sterols in vitro. The sterol binding by p33 is supported by cholesterol recognition/interaction amino acid consensus (CRAC) and CARC-like sequences within the two transmembrane domains of p33. Mutagenesis of the critical Y amino acids within the CRAC and CARC sequences blocked TBSV replication in yeast and plant cells. We also showed the enrichment of sterols in the detergent-resistant membrane (DRM) fractions obtained from yeast and plant cells replicating TBSV. The DRMs could support viral RNA synthesis on both the endogenous and exogenous templates. A lipidomic approach showed the lack of enhancement of sterol levels in yeast and plant cells replicating TBSV. The data support the notion that the TBSV replication proteins are associated with sterol-rich detergentresistant membranes in yeast and plant cells. Together, the results obtained in this study and the previously published results support the local enrichment of sterols around the viral replication proteins that is critical for TBSV replication.

Original languageEnglish
Article numbere01984-16
JournalJournal of Virology
Volume91
Issue number7
DOIs
StatePublished - 2017

Bibliographical note

Publisher Copyright:
© 2017 American Society for Microbiology. All Rights Reserved.

Keywords

  • Lipids
  • Membrane binding
  • Plant
  • RNA replication
  • Sterol
  • TBSV
  • Tombusvirus
  • Virus-host interactions
  • Yeast

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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