Regulation of peroxisomal proteins and organelle proliferation by multiple carbon sources in the methylotrophic yeast, Candida boidinii.
A methylotrophic yeast, Candida boidinii, was grown on various combinations of peroxisome-inducing carbon source(s) (PIC(s)), i.e. methanol, oleate and D-alanine, and the regulation of peroxisomal proteins (both matrix and membrane ones) and organelle proliferation were studied. This regulation was followed (1) at the protein or enzyme level by means of the peroxisomal enzyme activity and Western analysis; (2) at the mRNA level by Northern analysis; and (3) at the organelle level by direct observation of peroxisomes under a fluorescent microscope. Peroxisomal proliferation was followed in vivo by using a C. boidinii strain producing a green fluorescent protein having peroxisomal targeting signal 1. When multiple PICs were used for cell growth, C. boidinii induced specific peroxisomal proteins characteristic of all PIC(s) present in the medium, responding to all PIC(s) simultaneously. Thus, these PICs were considered to induce peroxisomal proliferation independently and not to repress peroxisomes induced by other PICs. Next, the sensitivity of the peroxisomal induction to glucose repression was studied. While the peroxisomal induction by methanol or oleate was completely repressed by glucose, the D-alanine-induced activities of D-amino acid oxidase and catalase, Pmp47, and the organelle proliferation were not. These results indicate that peroxisomal proliferation in yeasts is not necessarily sensitive to glucose repression. Lastly, this regulation was shown to occur at the mRNA level.
Sakai
Y
Y
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Japan. ysakai@kais.kyoto-u.ac.jp
Yurimoto
H
H
Matsuo
H
H
Kato
N
N
D042962:Acyl-CoA Oxidase; D000409:Alanine / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000032:analysis; D015152:Blotting, Northern; D015153:Blotting, Western; D002175:Candida / Q000235:genetics / Q000254:growth & development* / Q000648:ultrastructure; D002374:Catalase / Q000032:analysis; D003605:D-Amino-Acid Oxidase / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000378:metabolism; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents / Q000032:analysis; D008164:Luminescent Proteins / Q000032:analysis; D000432:Methanol / Q000378:metabolism*; D008830:Microbodies / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D019301:Oleic Acid / Q000378:metabolism; D015388:Organelles / Q000235:genetics / Q000378:metabolism*; D010088:Oxidoreductases / Q000032:analysis; D016133:Polymerase Chain Reaction; D012331:RNA, Fungal / Q000737:chemistry
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D012331:RNA, Fungal; D049452:Green Fluorescent Proteins; D019301:Oleic Acid; D010088:Oxidoreductases; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D002374:Catalase; D042962:Acyl-CoA Oxidase; D003605:D-Amino-Acid Oxidase; D005947:Glucose; D000409:Alanine; D000432:Methanol