Abstract
Sugar oxidoreductases such as cellobiose dehydrogenase or pyranose oxidase are widespread enzymes among fungi, whose biological function is largely speculative. We investigated a similar gene family in the mushroom Agaricus meleagris and its expression under various conditions. Three genes (named pdh1, pdh2 and pdh3) putatively encoding pyranose dehydrogenases were isolated. All three genes displayed a conserved structure and organization, and the respective cDNAs contained ORFs translating into polypeptides of 602 or 600 amino acids. The N-terminal sections of all three genes encode putative signal peptides consistent with the enzymes extracellular secretion. We cultivated the fungus on different carbon sources and analyzed the mRNA levels of all three genes over a period of several weeks using real-time RT-PCR. The glyceraldehyde-3-phosphate dehydrogenase gene from A. meleagris was also isolated and served as reference gene. pdh2 and pdh3 are essentially transcribed constitutively, whereas pdh1 expression is upregulated upon exhaustion of the carbon source; pdh1 appears to be additionally regulated under conditions of oxygen limitation. These data are consistent with an assumed role in lignocellulose degradation.
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Acknowledgments
This work was supported by the Austrian Science Fund (FWF) (grant P16836-B11 to CKP), the Ministry of Education, Youth and Sports of the Czech Republic (grants LC545, 6-06-4 and Institutional Research Concept AV0Z50200510, PH and JV), the Austrian Exchange Office (Scientific-Technical Cooperation Austria—Czech Republic, grant 2006/11) and the Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning, the Swedish Research Council, the CF Lundströms Stiftelse, and the Carl Tryggers Stiftelse (CD). We thank M. Zamocky, Department of Chemistry, University of Natural Resources and Applied Life Sciences, Vienna, for help with the phylogenetic analysis, and K. Brunner, Department of Chemical Engineering, Technical University Vienna, for technical help with the real-time transcript analysis.
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Communicated by U. Kües.
R. Kittl and C. Sygmund contributed equally to this work.
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Kittl, R., Sygmund, C., Halada, P. et al. Molecular cloning of three pyranose dehydrogenase-encoding genes from Agaricus meleagris and analysis of their expression by real-time RT-PCR. Curr Genet 53, 117–127 (2008). https://doi.org/10.1007/s00294-007-0171-9
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DOI: https://doi.org/10.1007/s00294-007-0171-9