Sadhale P P, Woychik N A
Roche Institute of Molecular Biology, Nutley, New Jersey 07110.
Mol Cell Biol. 1994 Sep;14(9):6164-70. doi: 10.1128/mcb.14.9.6164-6170.1994.
We identified a partially sequenced Saccharomyces cerevisiae gene which encodes a protein related to the S. cerevisiae RNA polymerase II subunit, RPB7. Several lines of evidence suggest that this related gene, YKL1, encodes the RNA polymerase III subunit C25. C25, like RPB7, is present in submolar ratios, easily dissociates from the enzyme, is essential for cell growth and viability, but is not required in certain transcription assays in vitro. YKL1 has ABF-1 and PAC upstream sequences often present in RNA polymerase subunit genes. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis mobility of the YKL1 gene product is equivalent to that of the RNA polymerase III subunit C25. Finally, a C25 conditional mutant grown at the nonpermissive temperature synthesizes tRNA at reduced rates relative to 5.8S rRNA, a hallmark of all characterized RNA polymerase III mutants.
我们鉴定出一个部分测序的酿酒酵母基因,它编码一种与酿酒酵母RNA聚合酶II亚基RPB7相关的蛋白质。多条证据表明,这个相关基因YKL1编码RNA聚合酶III亚基C25。C25与RPB7一样,以亚摩尔比例存在,很容易从酶上解离,对细胞生长和活力至关重要,但在某些体外转录试验中并非必需。YKL1具有ABF-1和PAC上游序列,这些序列常在RNA聚合酶亚基基因中出现。YKL1基因产物的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳迁移率与RNA聚合酶III亚基C25的迁移率相当。最后,在非允许温度下生长的C25条件突变体相对于5.8S rRNA以较低速率合成tRNA,这是所有已鉴定的RNA聚合酶III突变体的一个特征。