Ubiquitination of tombusvirus p33 replication protein plays a role in virus replication and binding to the host Vps23p ESCRT protein

Daniel Barajas, Peter D. Nagy

Research output: Contribution to journalArticlepeer-review

74 Scopus citations

Abstract

Post-translational modifications of viral replication proteins could be widespread phenomena during the replication of plus-stranded RNA viruses. In this article, we identify two lysines in the tombusvirus p33 replication co-factor involved in ubiquitination and show that the same lysines are also important for the p33 to interact with the host Vps23p ESCRT-I factor. We find that the interaction of p33 with Vps23p is also affected by a "late-domain"-like sequence in p33. The combined mutations of the two lysines and the late-domain-like sequences in p33 reduced replication of a replicon RNA of Tomato bushy stunt virus in yeast model host, in plant protoplasts, and plant leaves, suggesting that p33-Vps23p ESCRT protein interaction affects tombusvirus replication. Using ubiquitin-mimicking p33 chimeras, we demonstrate that high level of p33 ubiquitination is inhibitory for TBSV replication. These findings argue that optimal level of p33 ubiquitination plays a regulatory role during tombusvirus infections.

Original languageEnglish
Pages (from-to)358-368
Number of pages11
JournalVirology
Volume397
Issue number2
DOIs
StatePublished - Feb 20 2010

Bibliographical note

Funding Information:
We thank Drs. Judit Pogany and Zhenghe Li for critical reading of the manuscript and for their very helpful suggestions. The authors thank Dr. Scott D. Emr, Cornell University, for the yeast strains and plasmids. This work was supported by NIH-NIAID (5R21AI072170-02) and by the Kentucky Tobacco Research and Development Center at the University of Kentucky , awarded to P.D.N. D. Barajas was supported in part by a postdoctoral fellowship from the Spanish Ministry of Education and Science.

Keywords

  • ESCRT
  • Nicotiana benthamiana
  • RNA
  • Replication
  • Tobacco
  • Tomato bushy stunt virus
  • Ubiquitin
  • Yeast

ASJC Scopus subject areas

  • Virology

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