Bulawa C E
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.
Mol Cell Biol. 1992 Apr;12(4):1764-76. doi: 10.1128/mcb.12.4.1764-1776.1992.
In Saccharomyces cerevisiae, chitin forms the primary division septum and the bud scar in the walls of vegetative cells. Three chitin synthetic activities have been detected. Two of them, chitin synthase I and chitin synthase II, are not required for synthesis of most of the chitin present in vivo. Using a novel screen, I have identified three mutations, designated csd2, csd3, and csd4, that reduce levels of chitin in vivo by as much as 10-fold without causing any obvious perturbation of cell division. The csd2 and csd4 mutants lack chitin synthase III activity in vitro, while csd3 mutants have wild-type levels of this enzyme. In certain genetic backgrounds, these mutations cause temperature-sensitive growth on rich medium; inclusion of salts or sorbitol bypasses this phenotype. Gene disruption experiments show that CSD2 is nonessential; a small amount of chitin, about 5% of the wild-type level, is detected in the disruptants. DNA sequencing indicates that the CSD2 protein has limited, but statistically significant, similarity to chitin synthase I and chitin synthase II. Other significant similarities are to two developmental proteins: the nodC protein from Rhizobium species and the DG42 protein of Xenopus laevis. The relationship between the nodC and CSD2 proteins suggests that nodC may encode an N-acetylglucosaminyltransferase that synthesizes the oligosaccharide backbone of the nodulation factor NodRm-1.
在酿酒酵母中,几丁质形成了营养细胞细胞壁中的初级分裂隔膜和芽痕。已检测到三种几丁质合成活性。其中两种,几丁质合酶I和几丁质合酶II,对于体内大多数几丁质的合成并非必需。通过一种新颖的筛选方法,我鉴定出了三个突变,分别命名为csd2、csd3和csd4,它们可使体内几丁质水平降低多达10倍,且不会引起细胞分裂的任何明显紊乱。csd2和csd4突变体在体外缺乏几丁质合酶III活性,而csd3突变体具有该酶的野生型水平。在某些遗传背景下,这些突变会导致在丰富培养基上温度敏感型生长;添加盐或山梨醇可绕过这种表型。基因破坏实验表明CSD2并非必需;在破坏株中检测到少量几丁质,约为野生型水平的5%。DNA测序表明,CSD2蛋白与几丁质合酶I和几丁质合酶II具有有限但具有统计学意义的相似性。其他显著的相似性存在于两种发育蛋白中:来自根瘤菌属的nodC蛋白和非洲爪蟾的DG42蛋白。nodC和CSD2蛋白之间的关系表明,nodC可能编码一种合成结瘤因子NodRm-1寡糖主链的N-乙酰葡糖胺基转移酶。