Munro C A, Schofield D A, Gooday G W, Gow N A R
Department of Molecular & Cell Biology, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB25 2ZD, UK.
Microbiology (Reading). 1998 Feb;144 ( Pt 2):391-401. doi: 10.1099/00221287-144-2-391.
Candida albicans has three genes encoding chitin synthase enzymes. In wild-type strains, the expression of CHS2 and CHS3 peaked 1-2 h after the induction of hyphal growth, whilst mRNA levels in a non-germinative strain, CA2, remained low under the same conditions. CHS1 gene expression did not peak during germ tube formation but remained at low levels in both yeast and hyphal growth. The pattern of gene expression did not predict the changes in measured chitin synthase activities or changes in chitin content during dimorphic transition. Chitin synthase activity increased steadily, and did not peak shortly after germ tube induction, and activity profiles were similar in germ-tube-competent and germ-tube-negative strains. The phenotype of a delta chs2 null mutant suggested that CHS2 encoded the major enzyme activity in vitro and was largely responsible for elevated chitin synthase activities in microsomal preparations from hyphal cells compared to yeast cells. However, CaChs3p was responsible for synthesis of most chitin in both yeast and hyphae. Three independent chitin assays gave markedly different estimates of the relative chitin content of yeast and hyphae and wild-type and chs mutants. Only one of the methods gave a significantly higher chitin content for hyphal compared to yeast cell walls and a lower chitin content for hyphae of the delta chs2 null mutant compared to the parental strain.
白色念珠菌有三个编码几丁质合成酶的基因。在野生型菌株中,CHS2和CHS3的表达在菌丝生长诱导后1 - 2小时达到峰值,而在非发芽菌株CA2中,在相同条件下mRNA水平保持较低。CHS1基因表达在芽管形成期间未达到峰值,而是在酵母和菌丝生长过程中均保持在低水平。基因表达模式并未预测双态转变过程中几丁质合成酶活性的变化或几丁质含量的变化。几丁质合成酶活性稳步增加,在芽管诱导后不久未达到峰值,并且在有芽管形成能力的菌株和无芽管形成能力的菌株中活性曲线相似。Δchs2缺失突变体的表型表明,CHS2在体外编码主要的酶活性,并且与酵母细胞相比,在很大程度上负责菌丝体细胞微粒体制剂中几丁质合成酶活性的升高。然而,CaChs3p负责酵母和菌丝中大多数几丁质的合成。三种独立的几丁质测定方法对酵母和菌丝以及野生型和chs突变体的相对几丁质含量给出了明显不同的估计。只有一种方法显示,与酵母细胞壁相比,菌丝的几丁质含量显著更高,与亲本菌株相比,Δchs2缺失突变体菌丝的几丁质含量更低。