Hockman D J, Schultz M C
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Mol Cell Biol. 1996 Mar;16(3):892-8. doi: 10.1128/MCB.16.3.892.
Casein kinase II (CKII) is a ubiquitous and highly conserved serine/threonine protein kinase found in the nucleus and cytoplasm of most cells. Using a combined biochemical and genetic approach in the yeast Saccharomyces cerevisiae, we assessed the role of CKII in specific transcription by RNA polymerases I, II, and III. CKII is not required for basal transcription by RNA polymerases I and II but is important for polymerase III transcription. Polymerase III transcription is high in extracts with normal CKII activity but low in extracts from a temperature-sensitive mutant that has decreased CKII activity due to a lesion in the enzyme's catalytic alpha' subunit. Polymerase III transcription of 5S rRNA and tRNA templates in the temperature-sensitive extract is rescued by purified, wild-type CKII. An inhibitor of CKII represses polymerase III transcription in wild-type extract, and this repression is partly overcome by supplementing reaction mixtures with active CKII. Finally, we show that polymerase III transcription in vivo is impaired when CKII is inactivated. Our results demonstrate that CKII, an oncogenic protein kinase previously implicated in cell cycle and growth control, is required for high-level transcription by RNA polymerase III.
酪蛋白激酶II(CKII)是一种普遍存在且高度保守的丝氨酸/苏氨酸蛋白激酶,存在于大多数细胞的细胞核和细胞质中。我们在酿酒酵母中采用生物化学和遗传学相结合的方法,评估了CKII在RNA聚合酶I、II和III特异性转录中的作用。RNA聚合酶I和II的基础转录不需要CKII,但它对聚合酶III转录很重要。在具有正常CKII活性的提取物中,聚合酶III转录水平较高,而在一个温度敏感突变体的提取物中则较低,该突变体由于酶的催化α'亚基发生损伤而导致CKII活性降低。温度敏感提取物中5S rRNA和tRNA模板的聚合酶III转录可被纯化的野生型CKII挽救。CKII的一种抑制剂可抑制野生型提取物中的聚合酶III转录,通过在反应混合物中补充活性CKII可部分克服这种抑制作用。最后,我们表明当CKII失活时,体内的聚合酶III转录会受损。我们的结果表明,CKII这种先前与细胞周期和生长控制有关的致癌蛋白激酶,是RNA聚合酶III进行高水平转录所必需的。