Sudoh Masayuki, Tatsuno Kenji, Ono Naomi, Ohta Atsunori, Chibana Hiroji, Yamada-Okabe Hisafumi, Arisawa Mikio
Department of Mycology, Nippon Roche Research Center, 200 Kajiwara, Kamakura-shi, Kanagawa 247-8530, Japan.
Department of Genetics and Cell Biology, University of Minnesota, St Paul, Minnesota 55108-1095, USA.
Microbiology (Reading). 1999 Jul;145 ( Pt 7):1613-1622. doi: 10.1099/13500872-145-7-1613.
The Candida albicans CHS4 gene encoding chitin synthase 4 has been isolated using the Saccharomyces cerevisiae CHS4/SKT5 gene as a probe. The gene contains a 2061 bp open reading frame capable of encoding a protein of 687 amino acids (76053 Da). No intron was observed in the gene. Disruption of CHS4 in C. albicans yielded a Calcofluor-resistant phenotype, indicating that Chs4p contributes to chitin biosynthesis. Consistent with this, overexpression of Chs4p under the regulation of the ScGAL1 promoter enhanced chitin synthase 3 activity in S. cerevisiae 7- to 38-fold. In addition, chs3 and chs4 null mutants were significantly defective in Calcofluor white staining and their chitin content was 10% of that of the parental strain. Chs4p of C. albicans and S. cerevisiae showed 61% identity in the C-terminal half of the proteins and that region of C. albicans Chs4p complemented the Chs4p function of a mutant of S. cerevisiae resistant to Calcofluor white. Therefore, it appears that Chs4p is involved in chitin synthase 3 activity by combining with Chs3p to interact synergistically in chitin biosynthesis.
已使用酿酒酵母CHS4/SKT5基因作为探针分离出编码几丁质合成酶4的白色念珠菌CHS4基因。该基因包含一个2061 bp的开放阅读框,能够编码一个由687个氨基酸组成的蛋白质(76053 Da)。在该基因中未观察到内含子。白色念珠菌中CHS4的破坏产生了对荧光增白剂有抗性的表型,表明Chs4p有助于几丁质生物合成。与此一致的是,在ScGAL1启动子的调控下Chs4p的过表达使酿酒酵母中的几丁质合成酶3活性提高了7至38倍。此外,chs3和chs4缺失突变体在荧光增白剂染色方面存在显著缺陷,它们的几丁质含量是亲本菌株的10%。白色念珠菌和酿酒酵母的Chs4p在蛋白质的C端一半区域显示出61%的同一性,并且白色念珠菌Chs4p的该区域补充了对荧光增白剂有抗性的酿酒酵母突变体的Chs4p功能。因此,Chs4p似乎通过与Chs3p结合在几丁质生物合成中协同相互作用而参与几丁质合成酶3的活性。