Ohya Y, Anraku Y
Department of Biology, Faculty of Science, University of Tokyo, Japan.
Biochem Biophys Res Commun. 1989 Jan 31;158(2):541-7. doi: 10.1016/s0006-291x(89)80083-x.
The coding region of a chicken calmodulin cDNA was fused to a galactose-inducible GAL1 promoter, and an expression system was constructed in the yeast Saccharomyces cerevisiae. Expression of calmodulin was demonstrated by purifying the heterologously expressed protein and analyzing its biochemical properties. When the expression plasmid was introduced into a calmodulin gene (cmd1)-disrupted strain of yeast, the cells grew in galactose medium, showing that chicken calmodulin could complement the lesion of yeast calmodulin functionally. Repression of chicken calmodulin in the (cmd1)-disrupted strain caused cell cycle arrest with a G2/M nucleus, as observed previously with a conditional-lethal mutant of yeast calmodulin. These results suggest that the essential function of calmodulin for cell proliferation is conserved in cells ranging from yeast to vertebrate cells.
将鸡钙调蛋白cDNA的编码区与半乳糖诱导型GAL1启动子融合，并在酿酒酵母中构建了一个表达系统。通过纯化异源表达的蛋白并分析其生化特性，证实了钙调蛋白的表达。当将表达质粒导入酵母钙调蛋白基因（cmd1）缺失的菌株时，细胞能在半乳糖培养基中生长，表明鸡钙调蛋白在功能上可以弥补酵母钙调蛋白的缺陷。如之前在酵母钙调蛋白的条件致死突变体中观察到的那样，在（cmd1）缺失的菌株中抑制鸡钙调蛋白会导致细胞周期停滞在G2/M期细胞核。这些结果表明，从酵母到脊椎动物细胞，钙调蛋白对细胞增殖的基本功能是保守的。
