Sakai A, Shimizu Y, Kondou S, Chibazakura T, Hishinuma F
Laboratory of Molecular Genetics, Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Mol Cell Biol. 1990 Aug;10(8):4130-8. doi: 10.1128/mcb.10.8.4130-4138.1990.
An RGR1 gene product is required to repress expression of glucose-regulated genes in Saccharomyces cerevisiae. The abnormal morphology of rgr1 cells was studied. Scanning and transmission electron microscopic observations revealed that the cell wall of the daughter cell remained attached to that of mother cell. We cloned the RGR1 gene by complementation and showed that the cloned DNA was tightly linked to the chromosomal RGR1 locus. The cloned RGR1 gene suppressed all of the phenotypes caused by the mutation and encoded a 3.6-kilobase poly(A)+ RNA. The RGR1 gene is located on chromosome XII, as determined by pulsed-field gel electrophoresis, and we mapped rgr1 between gal2 and pep3 by genetic analysis. rgr1 was shown to be a new locus. We also determined the nucleotide sequence of RGR1, which was predicted to encode a 123-kilodalton protein. The null mutation resulted in lethality, indicating that the RGR1 gene is essential for growth. On the other hand, a carboxy-terminal deletion of the gene caused phenotypes similar to but more severe than those caused by the original mutation. The amount of reserve carbohydrates was reduced in rgr1 cells. Possible functions of the RGR1 product are discussed.
酿酒酵母中抑制葡萄糖调节基因的表达需要RGR1基因产物。对rgr1细胞的异常形态进行了研究。扫描电子显微镜和透射电子显微镜观察显示,子细胞的细胞壁与母细胞的细胞壁相连。我们通过互补克隆了RGR1基因,并表明克隆的DNA与染色体RGR1位点紧密相连。克隆的RGR1基因抑制了由该突变引起的所有表型,并编码一种3.6千碱基的聚腺苷酸加RNA。通过脉冲场凝胶电泳确定,RGR1基因位于第十二号染色体上,并且我们通过遗传分析将rgr1定位在gal2和pep3之间。rgr1被证明是一个新的位点。我们还确定了RGR1的核苷酸序列,预计它编码一种123千道尔顿的蛋白质。无效突变导致致死性,表明RGR1基因对生长至关重要。另一方面,该基因的羧基末端缺失导致的表型与原始突变相似,但更为严重。rgr1细胞中的储备碳水化合物含量降低。讨论了RGR1产物的可能功能。