RESUMO
The yeast gene, GRC5 (growth control), is a member of the highly conserved QM gene family, the human member of which has been associated with the suppression of Wilms' tumor. GRC5 encodes ribosomal protein L10, which is thought to play a regulatory role in the translational control of gene expression. A revertant screen identified four spontaneous revertants of the mutant grc5-1ts allele. Genetic and phenotypic analysis showed that these represent one gene, NMD3, and that the interaction of NMD3 and GRC5 is gene-specific. NMD3 was previously identified as a component of the nonsense-mediated mRNA decay pathway. The point mutations within NMD3 reported here may define a domain important for the functional interaction of Grc5p and Nmd3p.
Assuntos
Regulação da Expressão Gênica , Genes Fúngicos , Biossíntese de Proteínas , Saccharomyces cerevisiae/genética , Alelos , Sequência de Aminoácidos , Animais , Sequência de Bases , Primers do DNA/genética , Humanos , Dados de Sequência Molecular , Mutação , Fenótipo , Proteína Ribossômica L10 , Proteínas Ribossômicas/genética , Homologia de Sequência de Aminoácidos , Tumor de Wilms/genéticaRESUMO
We isolated the Saccharomyces cerevisiae GRC5 (growth control) gene by functional complementation in vivo of a ts (temperature sensitive) mutation. Phenotypic analysis suggested involvement of GRC5 in cell growth and proliferation. Mutant cells arrest their cell cycles after one to three cell divisions predominantly as mother cells with a large bud. In the region of the septum, a massive accumulation of cell wall material is observed. The mother and daughter nuclei are well separated and spindles are no longer present, while the cytoskeleton is of aberrant appearance. Arrested cells do not perform protein synthesis and are unable to mate. Furthermore, grc5-1ts cells rapidly lose viability at the restrictive temperature (37 degrees C) only on full media, but not under nitrogen-starvation conditions, indicating that proper response to this nutrient limitation is still intact in mutant cells after cell cycle arrest. The sequence of GRC5 translates into a basic protein of 221 amino acid with a corresponding Mr of 25.4 kDa. GRC5 is a member of the highly conserved QM gene family, members of which have been reported from plants, invertebrates and vertebrates. The amino acid sequence of GRC5 over its entire length is more than 60% identical to the human QM protein, expression of which is associated with loss of the tumorigenic phenotype in a cell line derived from Wilms' tumor, a malignancy of the embyronic kidney. Here, we show that GRC5 is an essential yeast gene, the function of which as inferred from analysis of the grc5-1ts mutant is crucial for establishment of proper cytoskeletal structure and regulation of growth in yeast cells.
