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A mutant allele of essential, general translation initiation factor DED1 selectively inhibits translation of a viral mRNA.

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A mutant allele of essential, general translation initiation factor DED1 selectively inhibits translation of a viral mRNA.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Chen, J. Noueiry, A. O. Ahlquist, P.
Description	Author Affiliation: Noueiry AO ( Institute for Molecular Virology, and Howard Hughes Medical Institute, University of Wisconsin, Madison, WI 53706, USA.);
Abstract	Positive-strand RNA virus genomes are substrates for translation, RNA replication, and encapsidation. To identify host factors involved in these functions, we used the ability of brome mosaic virus (BMV) RNA to replicate in yeast. We report herein identification of a mutation in the essential yeast gene DED1 that inhibited BMV RNA replication but not yeast growth. DED1 encodes a DEAD (Asp-Glu-Ala-Asp)-box RNA helicase required for translation initiation of all yeast mRNAs. Inhibition of BMV RNA replication by the mutant DED1 allele (ded1-18) resulted from inhibited expression of viral polymerase-like protein 2a, encoded by BMV RNA2. Inhibition of RNA2 translation was selective, with no effect on general cellular translation or translation of BMV RNA1-encoded replication factor 1a, and was independent of p20, a cellular antagonist of DED1 function in translation. Inhibition of RNA2 translation in ded1-18 yeast required the RNA2 5' noncoding region (NCR), which also conferred a ded1-18-specific reduction in expression on a reporter gene mRNA. Comparison of the similar RNA1 and RNA2 5' NCRs identified a 31-nucleotide RNA2-specific region that was required for the ded1-18-specific RNA2 translation block and attenuated RNA2 translation in wild-type yeast. Further comparisons and RNA structure predictions suggest a modular arrangement of replication and translation signals in RNA1 and RNA2 5' NCRs that appears conserved among bromoviruses. The 5' attenuator and DED1 dependence of RNA2 suggest that, despite its divided genome, BMV regulates polymerase translation relative to other replication factors, just as many single-component RNA viruses use translational read-through and frameshift mechanisms to down-regulate polymerase. The results show that a DEAD-box helicase can selectively activate translation of a specific mRNA and may provide a paradigm for translational regulation by other members of the ubiquitous DEAD-box RNA helicase family.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	24
Volume Number	97
Language	English
Publisher	National Academy of Sciences
Publisher Date	2000-12-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Bromovirus Genetics Cell Cycle Proteins Fungal Proteins Metabolism Gene Expression Regulation, Viral Protein Biosynthesis RNA Helicases RNA, Messenger RNA, Viral Saccharomyces Cerevisiae Proteins Virus Replication Physiology DEAD-box RNA Helicases Molecular Sequence Data Nucleic Acid Conformation Recombinant Proteins Biosynthesis Saccharomyces Cerevisiae Transcription, Genetic Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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