The Candida albicans CHS4 gene complements a Saccharomyces cerevisiae skt5/chs4 mutation and is involved in chitin biosynthesis.
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.
Sudoh
Masayuki
M
Department of Mycology, Nippon Roche Research Center, 200 Kajiwara, Kamakura-shi, Kanagawa 247-8530, Japan.
Tatsuno
Kenji
K
Department of Mycology, Nippon Roche Research Center, 200 Kajiwara, Kamakura-shi, Kanagawa 247-8530, Japan.
Ono
Naomi
N
Department of Mycology, Nippon Roche Research Center, 200 Kajiwara, Kamakura-shi, Kanagawa 247-8530, Japan.
Ohta
Atsunori
A
Department of Mycology, Nippon Roche Research Center, 200 Kajiwara, Kamakura-shi, Kanagawa 247-8530, Japan.
Chibana
Hiroji
H
Department of Genetics and Cell Biology, University of Minnesota, St Paul, Minnesota 55108-1095, USA.
Yamada-Okabe
Hisafumi
H
Department of Mycology, Nippon Roche Research Center, 200 Kajiwara, Kamakura-shi, Kanagawa 247-8530, Japan.
Arisawa
Mikio
M
Department of Mycology, Nippon Roche Research Center, 200 Kajiwara, Kamakura-shi, Kanagawa 247-8530, Japan.
D000595:Amino Acid Sequence; D001557:Benzenesulfonates; D015139:Blotting, Southern; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure; D002473:Cell Wall / Q000737:chemistry; D002686:Chitin / Q000032:analysis / Q000096:biosynthesis*; D002687:Chitin Synthase / Q000235:genetics* / Q000378:metabolism*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment; D013194:Staining and Labeling
D001557:Benzenesulfonates; D002686:Chitin; C007061:C.I. Fluorescent Brightening Agent 28; D002687:Chitin Synthase