title: Unique C-terminal region of Hap3 is required for methanol-regulated gene expression in the methylotrophic yeast Candida boidinii.
abstract: The Hap complex of the methylotrophic yeast Candida boidinii was found to be required for methanol-regulated gene expression. In this study, we performed functional characterization of CbHap3p, one of the Hap complex components in C. boidinii. Sequence alignment of Hap3 proteins revealed the presence of a unique extended C-terminal region, which is not present in Hap3p from Saccharomyces cerevisiae (ScHap3p), but is found in Hap3p proteins of methylotrophic yeasts. Deletion of the C-terminal region of CbHap3p (Δ256-292 or Δ107-237) diminished activation of methanol-regulated genes and abolished the ability to grow on methanol, but did not affect nuclear localization or DNA-binding ability. However, deletion of the N-terminal region of CbHap3p (Δ1-20) led to not only a growth defect on methanol and a decreased level of methanol-regulated gene expression, but also impaired nuclear localization and binding to methanol-regulated gene promoters. We also revealed that CbHap3p could complement the growth defect of the Schap3Δ strain on glycerol, although ScHap3p could not complement the growth defect of a Cbhap3Δ strain on methanol. We conclude that the unique C-terminal region of CbHap3p contributes to maximum activation of methanol-regulated genes, whilst the N-terminal region is required for nuclear localization and binding to DNA.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University,Kyoto,Japan.
authors.forename: Saori
authors.initials: S
authors.lastname: Oda
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University,Kyoto,Japan.
authors.forename: Hiroya
authors.initials: H
authors.lastname: Yurimoto
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University,Kyoto,Japan.
authors.forename: Nobuhisa
authors.initials: N
authors.lastname: Nitta
authors.affiliation: Research Unit for Physiological Chemistry, Center for the Promotion of Interdisciplinary Education and Research, Kyoto University,Kyoto,Japan.|Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University,Kyoto,Japan.
authors.forename: Yasuyoshi
authors.initials: Y
authors.lastname: Sakai
mesh_terms: D000595:Amino Acid Sequence; D023081:CCAAT-Binding Factor / Q000235:genetics*; D002175:Candida / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics; D015870:Gene Expression / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D005990:Glycerol / Q000378:metabolism; D000432:Methanol / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*
chemical_list: D023081:CCAAT-Binding Factor; D004268:DNA-Binding Proteins; C490431:HAP3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D005990:Glycerol; D000432:Methanol