Published online 2 April 2004
Nucleic Acids Research, 2004, Vol. 32, No. 6 2031-2038
© 2004 Oxford University Press
Ded1p, a conserved DExD/H-box translation factor, can promote yeast L-A virus negative-strand RNA synthesis in vitro
1 Department of Molecular Genetics and 2 Program of Molecular, Cellular and Developmental Biology, The Ohio State University, Columbus, OH 43210, USA
*To whom correspondence should be addressed. Tel: +1 614 688 8678; Fax: +1 614 292 4466; Email: chang.108{at}osu.edu
Present address:
Ray-Yuan Chuang, Institute for Biological Energy Alternatives, 9700 Great Seneca Highway, Laboratory 174, Rockville, MD 20850, USA
Received February 18, 2004; Revised and Accepted March 10, 2004
Viruses are intracellular parasites that must use the host machinery to multiply. Identification of the host factors that perform essential functions in viral replication is thus of crucial importance to the understanding of virus–host interactions. Here we describe Ded1p, a highly conserved DExD/H-box translation factor, as a possible host factor recruited by the yeast L-A double-stranded RNA (dsRNA) virus. We found that Ded1p interacts specifically and strongly with Gag, the L-A virus coat protein. Further analysis revealed that Ded1p interacts with the L-A virus in an RNA-independent manner and, as a result, L-A particles can be affinity purified via this interaction. The affinity-purified L-A particles are functional, as they are capable of synthesizing RNA in vitro. Critically, using purified L-A particles, we demonstrated that Ded1p specifically promotes L-A dsRNA replication by accelerating the rate of negative-strand RNA synthesis in vitro. In light of these data, we suggest that Ded1p may be a part of the long sought after activity shown to promote yeast viral dsRNA replication. This and the fact that Ded1p is also required for translating brome mosaic virus RNA2 in yeast thus raise the intriguing possibility that Ded1p is one of the key host factors favored by several evolutionarily related RNA viruses, including the human hepatitis C virus.
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