Sakurai H, Kimura M, Ishihama A
National Institute of Genetics, Department of Molecular Genetics, Shizuoka, Japan.
Gene. 1998 Oct 9;221(1):11-6. doi: 10.1016/s0378-1119(98)00449-1.
Both the rpb9 gene and its cDNA encoding the subunit 9 of RNA polymerase II were cloned from the fission yeast Schizosaccharomyces pombe. From the DNA sequences, Rpb9 was predicted to consist of 113 amino acid residues with a molecular mass of 13,175. S. pombe Rpb9 is 47, 40 and 36% identical in amino acid sequence to the corresponding subunits from Saccharomyces cerevisiae, human and Drosophila melanogaster, respectively. Previously, we failed to detect Rpb9 in the purified RNA polymerase II by amino-terminal micro-sequencing of proteolytic fragments of subunits separated by SDS-gel electrophoresis. After Western blot analysis using antibodies raised against the protein product of the newly isolated rpb9 gene, we found that the purified RNA polymerase II contains Rpb9.
RNA聚合酶II亚基9的编码基因rpb9及其cDNA均从裂殖酵母粟酒裂殖酵母中克隆得到。根据DNA序列预测,Rpb9由113个氨基酸残基组成,分子量为13175。粟酒裂殖酵母Rpb9的氨基酸序列与酿酒酵母、人类和黑腹果蝇相应亚基的氨基酸序列分别有47%、40%和36%的同一性。此前,我们通过SDS凝胶电泳分离的亚基蛋白水解片段的氨基末端微测序,未能在纯化的RNA聚合酶II中检测到Rpb9。在用针对新分离的rpb9基因的蛋白质产物产生的抗体进行蛋白质印迹分析后,我们发现纯化的RNA聚合酶II含有Rpb9。