Translation elongation factor 1A is a component of the tombusvirus replicase complex and affects the stability of the p33 replication co-factor

Zhenghe Li, Judit Pogany, Tadas Panavas, Kai Xu, Anthony M. Esposito, Terri Goss Kinzy, Peter D. Nagy

Research output: Contribution to journalArticlepeer-review

116 Scopus citations

Abstract

Host RNA-binding proteins are likely to play multiple, integral roles during replication of plus-strand RNA viruses. To identify host proteins that bind to viral RNAs, we took a global approach based on the yeast proteome microarray, which contains 4080 purified yeast proteins. The biotin-labeled RNA probes included two distantly related RNA viruses, namely Tomato bushy stunt virus (TBSV) and Brome mosaic virus (BMV). Altogether, we have identified 57 yeast proteins that bound to TBSV RNA and/or BMV RNA. Among the identified host proteins, eleven bound to TBSV RNA and seven bound to BMV RNA with high selectivity, whereas the remaining 39 host proteins bound to both viral RNAs. The interaction between the TBSV replicon RNA and five of the identified host proteins was confirmed via gel-mobility shift and co-purification experiments from yeast. Over-expression of the host proteins in yeast, a model host for TBSV, revealed 4 host proteins that enhanced TBSV replication as well as 14 proteins that inhibited replication. Detailed analysis of one of the identified yeast proteins binding to TBSV RNA, namely translation elongation factor eEF1A, revealed that it is present in the highly purified tombusvirus replicase complex. We also demonstrate binding of eEF1A to the p33 replication protein and a known cis-acting element at the 3′ end of TBSV RNA. Using a functional mutant of eEF1A, we provide evidence on the involvement of eEF1A in TBSV replication.

Original languageEnglish
Pages (from-to)245-260
Number of pages16
JournalVirology
Volume385
Issue number1
DOIs
StatePublished - Mar 1 2009

Bibliographical note

Funding Information:
The authors thank Drs. David Smith and Daniel Barajas for valuable comments. This work was supported by the National Science Foundation (IOB-0517218), NIH-NIAID and the Kentucky Tobacco Research and Development Center to PDN, NIH T32AI007403 (AME) and NIH RO1GM57483 (TGK).

Keywords

  • Host factor
  • Protein array
  • Proteome array
  • Replicase
  • Replication
  • TEF1
  • Tomato bushy stunt virus
  • Yeast as a model host
  • eEF1A

ASJC Scopus subject areas

  • Virology

Fingerprint

Dive into the research topics of 'Translation elongation factor 1A is a component of the tombusvirus replicase complex and affects the stability of the p33 replication co-factor'. Together they form a unique fingerprint.

Cite this