Control of translational repression by protein-protein interactions.
The coat protein of the RNA bacteriophage MS2 is a translational repressor and interacts with a specific RNA stem-loop to inhibit translation of the viral replicase gene. As part of an effort to dissect genetically its RNA binding function, mutations were identified in the coat protein sequence that suppress mutational defects in the translational operator. Each of the mutants displayed a super-repressor phenotype, repressing translation from the wild-type and a variety of mutant operators better than did the wild-type coat protein. At least one mutant probably binds RNA more tightly than wild-type. The other mutants, however, were defective for assembly of virus-like particles, and self-associated predominantly as dimers. It is proposed that this assembly defect accounts for their super-repressor characteristics, since failure to assemble into virus-like particles elevates the effective concentration of repressor dimers. This hypothesis is supported by the observation that deletion of thirteen amino acids known to be important for assembly of dimers into capsids also resulted in the same assembly defect and in super-repressor activity. A second class of assembly defects is also described. Deletion of two amino acids from the C-terminus of coat protein resulted in failure to form capsids, most of the coat protein having the apparent molecular weight expected of trimers. This mutant (dl-8) was completely defective for repressor activity, probably because of an inability to form dimers. These results point out the inter-dependence of the structural and regulatory functions of coat protein.
Peabody
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Department of Cell Biology, University of New Mexico School of Medicine, Albuquerque 87131.
Ely
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D000595:Amino Acid Sequence; D001483:Base Sequence; D002213:Capsid / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D036022:Capsid Proteins; D003090:Coliphages / Q000235:genetics* / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics*
D036022:Capsid Proteins; D046911:Macromolecular Substances; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D012324:RNA-Dependent RNA Polymerase