Molecular characterization of Candida boidinii MIG1 and its role in the regulation of methanol-inducible gene expression.
Methanol-inducible gene promoters in methanol-utilizing yeasts are used in high-level heterologous gene expression systems. Generally, expression of methanol-inducible genes is completely repressed by the presence of glucose. In this study we identified the MIG1 gene in Candida boidinii, which encodes a homologue of the glucose repressor Mig1p of Saccharomyces cerevisiae. Disruption of the CbMIG1 gene had no growth effect on various carbon sources. Activation of the methanol-inducible AOD1 gene, which encodes alcohol oxidase, was increased in the early stage of methanol induction when cells of the CbMIG1-disrupted strain were transferred from glucose medium to methanol medium. Furthermore, CbMig1p tagged with yellow fluorescent protein was primarily localized in the nucleus of glucose-grown cells, but was diffuse in the cytosol of methanol-grown cells. This cytosolic diffusion in methanol-grown cells occurred in a CbMsn5p-dependent manner. These results suggest that CbMig1p is involved in negative regulation of methanol-inducible gene expression in C. boidinii.
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Japan.
Zhenyu
Z
Zhai
Hiroya
H
Yurimoto
Yasuyoshi
Y
Sakai
D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D002175:Candida / Q000737:chemistry / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D002453:Cell Cycle; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000378:metabolism; D000432:Methanol / Q000378:metabolism*; D008969:Molecular Sequence Data; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment
D005656:Fungal Proteins; C085976:MIG1 protein, S cerevisiae; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D005947:Glucose; D000432:Methanol