Padmanabha R, Chen-Wu J L, Hanna D E, Glover C V
Department of Biochemistry, University of Georgia, Athens 30602.
Mol Cell Biol. 1990 Aug;10(8):4089-99. doi: 10.1128/mcb.10.8.4089-4099.1990.
Casein kinase II of Saccharomyces cerevisiae contains two distinct catalytic subunits, alpha and alpha', which are encoded by the CKA1 and CKA2 genes, respectively. Null mutations in the CKA1 gene do not confer a detectable phenotype (J. L.-P. Chen-Wu, R. Padmanabha, and C. V. C. Glover, Mol. Cell. Biol. 8:4981-4990, 1988), presumably because of the presence of the CKA2 gene. We report here the cloning, sequencing, and disruption of the CKA2 gene. The alpha' subunit encoded by the CKA2 gene is 60% identical to the CKA1-encoded alpha subunit and 55% identical to the Drosophila alpha subunit (A. Saxena, R. Padmanabha, and C. V. C. Glover, Mol. Cell. Biol. 7:3409-3417, 1987). Deletions of the CKA2 gene were constructed by gene replacement techniques. Haploid cells in which the CKA2 gene alone is disrupted show no detectable phenotype, but haploid cells carrying disruptions in both the CKA1 and CKA2 genes are inviable. Cells in which casein kinase II activity is depleted increase substantially in size prior to growth arrest, and a significant fraction of the arrested cells exhibit a pseudomycelial morphology. Disruption of the activity also results in flocculation. Yeast strains lacking both endogenous catalytic subunit genes can be rescued by expression of the alpha and beta subunits of Drosophila casein kinase II or by expression of the Drosophila alpha subunit alone, suggesting that casein kinase II function has been conserved through evolution.
酿酒酵母的酪蛋白激酶II包含两个不同的催化亚基,α和α',它们分别由CKA1和CKA2基因编码。CKA1基因的无效突变不会产生可检测到的表型(J. L.-P. Chen-Wu、R. Padmanabha和C. V. C. Glover,《分子细胞生物学》8:4981 - 4990,1988),推测是因为CKA2基因的存在。我们在此报告CKA2基因的克隆、测序和破坏。由CKA2基因编码的α'亚基与CKA1编码的α亚基有60%的同一性,与果蝇的α亚基有55%的同一性(A. Saxena、R. Padmanabha和C. V. C. Glover,《分子细胞生物学》7:3409 - 3417,1987)。通过基因替换技术构建了CKA2基因的缺失。仅CKA2基因被破坏的单倍体细胞没有可检测到的表型,但CKA1和CKA2基因都被破坏的单倍体细胞无法存活。酪蛋白激酶II活性被耗尽的细胞在生长停滞之前体积会大幅增加,并且很大一部分停滞的细胞呈现假菌丝形态。活性的破坏也会导致絮凝。缺乏两个内源性催化亚基基因的酵母菌株可以通过表达果蝇酪蛋白激酶II的α和β亚基或仅表达果蝇α亚基来挽救,这表明酪蛋白激酶II的功能在进化过程中得以保留。