Abstract
Recruited host factors aid replication of plus-strand RNA viruses. In this paper, we show that Dbp2 DEAD-box helicase of yeast, which is a homolog of human p68 DEAD-box helicase, directly affects replication of Tomato bushy stunt virus (TBSV). We demonstrate that Dbp2 binds to the 3'-end of the viral minus-stranded RNA and enhances plus-strand synthesis by the viral replicase in a yeast-based cell-free TBSV replication assay. In vitro data with wt and an ATPase-deficient Dbp2 mutant indicate that Dbp2 unwinds local secondary structures at the 3'-end of the TBSV (-)RNA. We also show that Dbp2 complements the replication deficiency of TBSV in yeast containing reduced amount of Ded1 DEAD-box helicase, another host factor involved in TBSV replication, suggesting that Dbp2 and Ded1 helicases play redundant roles in TBSV replication. We also show that the orthologous AtRH20 DEAD-box helicase from Arabidopsis can increase tombusvirus replication in vitro and in yeast.
Original language | English |
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Pages (from-to) | 470-484 |
Number of pages | 15 |
Journal | Virology |
Volume | 432 |
Issue number | 2 |
DOIs | |
State | Published - Oct 25 2012 |
Bibliographical note
Funding Information:The authors thank Dr. Judit Pogany for valuable comments. dbp2-F366L plasmid was the generous gift of Dr. N.K. Tanner. This work was supported by NIH-NIAID ( 5R21AI079457-02 and 5R21AI072170-02 ) and University of Kentucky.
Keywords
- Arabidopsis
- Cell-free assay
- DEAD-box helicase
- Host factor
- In vitro RNA synthesis
- RNA unwinding
- RNA virus replication
- Yeast
ASJC Scopus subject areas
- Virology