Miyakawa Y
Department of Life Sciences, Yamanashi Medical College, Tamaho, Yamanashi 409-3898, Japan.
Yeast. 2000 Aug;16(11):1045-51. doi: 10.1002/1097-0061(200008)16:11<1045::AID-YEA595>3.0.CO;2-L.
In a screen for the protein kinase genes of the human pathogenic yeast Candida albicans, a putative homologue (CaPHO85) of PHO85, a negative regulator of the PHO system of Saccharomyces cerevisiae, which is one of the cyclin-dependent protein kinases (CDKs), was isolated. An open reading frame (ORF) of this gene was identified encoding a predicted protein of 326 amino acids with a calculated molecular weight of 37.6 kDa. The amino acid sequence is highly homologous to S. cerevisiae Pho85 (62% identity) and its Aspergillus nidulans homologue (70% identity), but less homologous to Cdc28 (50% identity) of S. cerevisiae and to its C. albicans homologue CaCdc28 (49% identity), both of which are also CDK. The coding region for the C. albicans gene was interrupted by an intron of 81 nucleotides near the sequence encoding the N-terminal region, similarly to the case of the S. cerevisiae PHO85 gene. Alignment of CaPho85 with various yeast CDKs revealed that most of the domains for ATP-binding and protein kinase activity are conserved among fungal species. Southern blot analysis indicated that CaPHO85 is most likely present as a single copy gene. This gene complemented the pho85 mutation of S. cerevisiae by transformation.
在对人类致病酵母白色念珠菌的蛋白激酶基因进行筛选时,分离出了酿酒酵母PHO系统的负调控因子PHO85(一种细胞周期蛋白依赖性蛋白激酶(CDK))的一个假定同源物(CaPHO85)。鉴定出该基因的一个开放阅读框(ORF),其编码一个预测的由326个氨基酸组成的蛋白质,计算分子量为37.6 kDa。该氨基酸序列与酿酒酵母Pho85高度同源(同一性为62%)及其烟曲霉同源物(同一性为70%),但与酿酒酵母的Cdc28(同一性为50%)及其白色念珠菌同源物CaCdc28(同一性为49%)的同源性较低,这两者也都是CDK。白色念珠菌基因的编码区在编码N端区域的序列附近被一个81个核苷酸的内含子中断,这与酿酒酵母PHO85基因的情况类似。CaPho85与各种酵母CDK的比对显示,大多数ATP结合和蛋白激酶活性结构域在真菌物种中是保守的。Southern印迹分析表明,CaPHO85很可能以单拷贝基因形式存在。通过转化,该基因互补了酿酒酵母的pho85突变。