Bulawa C E, Slater M, Cabib E, Au-Young J, Sburlati A, Adair W L, Robbins P W
Cell. 1986 Jul 18;46(2):213-25. doi: 10.1016/0092-8674(86)90738-5.
The chitin synthase of Saccharomyces is a plasma membrane-bound zymogen. Following proteolytic activation, the enzyme synthesizes insoluble chitin that has chain length and other physical properties similar to chitin found in bud scars. We isolated mutants lacking chitin synthase activity (chs1) and used these to clone CHS1. The gene has an open reading frame of 3400 bases and encodes a protein of 130 kd. The fission yeast S. pombe lacks chitin synthase and chitin. When a plasmid encoding a CHS1-lacZ fusion protein is introduced into S. pombe, both enzymatic activities are expressed in the same ratio as in S. cerevisiae, demonstrating that CHS1 encodes the structural gene of chitin synthase. Three CHS1 gene disruption experiments were performed. In all cases, strains with the disrupted gene have a recognizable phenotype, lack measurable chitin synthase activity in vitro but are viable, contain normal levels of chitin in vivo, and mate and sporulate efficiently.
酿酒酵母的几丁质合酶是一种与质膜结合的酶原。经过蛋白水解激活后，该酶合成不溶性几丁质，其链长和其他物理性质与芽痕中发现的几丁质相似。我们分离出缺乏几丁质合酶活性的突变体（chs1），并利用这些突变体克隆了CHS1。该基因有一个3400个碱基的开放阅读框，编码一个130kd的蛋白质。裂殖酵母粟酒裂殖酵母缺乏几丁质合酶和几丁质。当将编码CHS1 - lacZ融合蛋白的质粒导入粟酒裂殖酵母时，两种酶活性的表达比例与酿酒酵母相同，这表明CHS1编码几丁质合酶的结构基因。进行了三次CHS1基因破坏实验。在所有情况下，基因被破坏的菌株都有可识别的表型，体外缺乏可测量的几丁质合酶活性，但仍可存活，体内几丁质水平正常，并且能够高效交配和形成孢子。
