Matsuo Yasuhiro, Tanaka Katsunori, Matsuda Hideyuki, Kawamukai Makoto
Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Matsue, Japan.
FEBS Lett. 2005 May 9;579(12):2737-43. doi: 10.1016/j.febslet.2005.04.008. Epub 2005 Apr 19.
In Schizosaccharomyces pombe, a major role of chitin is to build up a complete spore. Here, we analyzed the cda1(+) gene (SPAC19G12.03), which encodes a protein homologous to chitin deacetylases, to know whether it is required for spore formation in S. pombe. The homothallic Deltacda1 strain constructed by homologous recombination was found to form a little amount of abnormal spores that contained one, two, or three asci, similar to (but not as strong as) the phenotype observed in a deletion mutant of chs1 encoding chitin synthase 1. This phenotype is reversed by expression of S. cerevisiae chitin deacetylase CDA1 or CDA2, suggesting that cda1 encodes a chitin deacetylase. To support the role of Cda1 in sporulation, the timing of expression of cda1(+) mRNA increased during sporulation process. We also found that the Cda1 protein self-associated when its binding was tested both by two-hybrid system and immunoprecipitation. Thus, these data indicated that cda1(+) is required for proper spore formation in S. pombe.
在粟酒裂殖酵母中,几丁质的一个主要作用是构建完整的孢子。在此,我们分析了编码与几丁质脱乙酰酶同源蛋白的cda1(+)基因(SPAC19G12.03),以了解它是否是粟酒裂殖酵母孢子形成所必需的。通过同源重组构建的同宗配合Δcda1菌株形成了少量异常孢子,这些孢子含有一个、两个或三个子囊,类似于(但不如)在编码几丁质合酶1的chs1缺失突变体中观察到的表型。酿酒酵母几丁质脱乙酰酶CDA1或CDA2的表达可逆转此表型,这表明cda1编码一种几丁质脱乙酰酶。为了支持Cda1在孢子形成中的作用,cda1(+)mRNA的表达时间在孢子形成过程中增加。我们还发现,通过双杂交系统和免疫沉淀检测其结合时,Cda1蛋白会自我结合。因此,这些数据表明cda1(+)是粟酒裂殖酵母正常孢子形成所必需的。