Munro Carol A, Whitton Rhian K, Hughes H Bleddyn, Rella Monika, Selvaggini Serena, Gow Neil A R
Department of Molecular and Cell Biology, University of Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, UK.
Fungal Genet Biol. 2003 Nov;40(2):146-58. doi: 10.1016/s1087-1845(03)00083-5.
In silico analysis of the genome sequence of the human pathogenic fungus Candida albicans identified an open reading frame encoding a putative fourth member of the chitin synthase gene family. This gene, named CaCHS8, encodes an 1105 amino acid open reading frame with the conserved motifs characteristic of class I zymogenic chitin synthases with closest sequence similarity to the non-essential C. albicans class I CHS2 gene. Although the CaCHS8 gene was expressed in both yeast and hyphal cells, homozygous chs8 Delta null mutants had normal growth rates, cellular morphologies and chitin contents. The null mutant strains had a 25% reduction in chitin synthase activity and were hypersensitive to Calcofluor White. A chs2 Delta chs8 Delta double mutant had less than 3% of normal chitin synthase activity and had increased wall glucan and decreased mannan but was unaffected in growth or cell morphology. The C. albicans class I double mutant did not exhibit a bud-lysis phenotype as found in the class I chs1 Delta mutant of Saccharomyces cerevisiae. Therefore, C. albicans has four chitin synthases with two non-essential class I Chs isoenzymes that contribute collectively to more than 97% of the in vitro chitin synthase activity.
对人类致病真菌白色念珠菌的基因组序列进行的电子分析鉴定出一个开放阅读框,其编码几丁质合酶基因家族的一个假定的第四个成员。这个名为CaCHS8的基因编码一个由1105个氨基酸组成的开放阅读框,具有I类产酶原几丁质合酶的保守基序,与非必需的白色念珠菌I类CHS2基因的序列相似性最高。尽管CaCHS8基因在酵母细胞和菌丝细胞中均有表达,但纯合的chs8Δ缺失突变体具有正常的生长速率、细胞形态和几丁质含量。缺失突变体菌株的几丁质合酶活性降低了25%,并且对荧光增白剂高度敏感。chs2Δchs8Δ双突变体的几丁质合酶活性不到正常水平的3%,其细胞壁葡聚糖增加,甘露聚糖减少,但生长或细胞形态未受影响。白色念珠菌I类双突变体并未表现出酿酒酵母I类chs1Δ突变体中发现的芽裂解表型。因此,白色念珠菌有四种几丁质合酶,其中两种非必需的I类Chs同工酶共同贡献了体外几丁质合酶活性的97%以上。