The (+) strand of the L-A dsRNA virus of Saccharomyces cerevisiae has two large open reading frames, ORF1, encoding the major coat protein, and ORF2 that encodes a single-stranded RNA binding protein having a sequence diagnostic of viral RNA-dependent RNA polymerases. ORF2 is expressed only as a """"""""gagpol"""""""" type fusion protein with ORF1. We have constructed a plasmid expressing these proteins and show that it can support the replication of the killer toxin-encoding Ml satellite virus in the absence of an L-A dsRNA helper virus. ORF1 expression makes the strain a host ski-phenocopy, suppressing M1's requirement for MAK11, MAK18, and MAK27 genes, allowing a defective L-A (L-A-E) to support M1 replication and making a wild-type killer a superkiller. We have defined the packaging signal present on L-A (+) strands and on the (+) strands of satellite viruses of L-A. into When this signal is inserted foreign RNAs, those RNAs are packaged in L-A virus particles. have we demonstrated that ORF1 and ORF2 are fused to make the """"""""gag-pol"""""""" fusion protein by -1 ribosomal frameshifting, the same mechanism used by retroviruses for this purpose. Our studies of this process in the case of L-A show that the strength of mRNA-tRNA binding at the ribosomal A-site is an important factor determining the efficiency of frameshifting. we propose that the fusion proteins of L-A and retroviruses use their pol domain to hold onto genomic RNA while their gag domain associates with gag molecules to prime coat formation.

Project Start
Project End
Budget Start
Budget End
Support Year
17
Fiscal Year
1990
Total Cost
Indirect Cost
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Country
United States
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Tang, Jinghua; Naitow, Hisashi; Gardner, Nora A et al. (2005) The structural basis of recognition and removal of cellular mRNA 7-methyl G 'caps' by a viral capsid protein: a unique viral response to host defense. J Mol Recognit 18:158-68
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