Functional complementation of catalase-defective peroxisomes in a methylotrophic yeast by import of the catalase A from Saccharomyces cerevisiae.
A mutant of the methanol-utilizing yeast Hansenula polymorpha defective in catalase was isolated. It lacks the ability to grow on methanol as the sole source of carbon and energy due to a loss of peroxisomal function that is required for the dissimilation and assimilation of this substrate. Growth of the mutant on glucose or glycerol was not impaired. Transformation of mutant cells with the gene coding for catalase A from Saccharomyces cerevisiae [Cohen, G., Fessl, F., Traczyk, J., Rytka, J. & Ruis, H. (1985) Mol. Gen. Genet. 200, 74-79] conferred constitutive expression of catalase activity. When the gene was placed under control of the regulatory methanol oxidase promoter from H. polymorpha, high levels of activity subject to glucose repression were obtained. In both cases efficient targeting of catalase A to the heterologous peroxisomes and assembly into an active form could be demonstrated. Concomitantly, growth on methanol was restored in the transformed mutant. The results are in line with a high conservation of transport signals on peroxisomal proteins. Expression of a cytosolic catalase in H. polymorpha did not confer the ability to grow on methanol. Therefore, proper localization of the catalase activity is a prerequisite for peroxisomal function.
Hansen
H
H
Institut für Mikrobiologie der Heinrich-Heine-Universität Düsseldorf, Federal Republic of Germany.
Roggenkamp
R
R
D000429:Alcohol Oxidoreductases / Q000235:genetics; D002374:Catalase / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015151:Immunoblotting; D007700:Kinetics; D000432:Methanol / Q000378:metabolism*; D008830:Microbodies / Q000201:enzymology* / Q000648:ultrastructure; D009154:Mutation; D010843:Pichia / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D004718:Saccharomycetales / Q000235:genetics*; D013728:Terminator Regions, Genetic; D014170:Transformation, Genetic
D000429:Alcohol Oxidoreductases; C029293:methanol oxidase; D002374:Catalase; D000432:Methanol