Protease inhibitors clitocypin and macrocypin are differentially expressed within basidiomycete fruiting bodies |
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Authors: | Sabotič Jerica Kilaru Sreedhar Budič Maruška Gašparič Meti Buh Gruden Kristina Bailey Andy M Foster Gary D Kos Janko |
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Affiliation: | aDepartment of Biotechnology, Jo?ef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia;bSchool of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, United Kingdom;cDepartment of Biotechnology and Systems Biology, National Institute of Biology, Ve?na pot 111, 1000 Ljubljana, Slovenia |
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Abstract: | Clitocypin and macrocypin are cysteine protease inhibitors of the mycocypin family which is unique to basidiomycetes. We have established that Clitocybe nebularis and Macrolepiota procera each contain genes for both macrocypin and clitocypin. Both are expressed in M. procera but only clitocypin in C. nebularis. Further analysis of mycocypin expression at the mRNA and protein levels in mature fruiting bodies of M. procera revealed that clitocypin is expressed evenly throughout the fruiting body, while the level of expression of macrocypins varies, and, at the protein level, is much higher in the veil fragments and the ring. The expression patterns of various mycocypins were determined in Coprinopsis cinerea, using promoters linked to a reporter gene. The expression profile of the clitocypin promoter was similar to that of the constitutive promoter gpdII from Agaricus bisporus, while that of the macrocypin 4 promoter was limited to the outer edges of the fruiting body throughout development. In addition, the activity of the macrocypin 3 promoter was different, indicating different regulation of expression for different macrocypin genes. The complex, tissue specific expression patterns for mycocypin genes suggest different biological roles for the products, either in regulation of endogenous proteases or in defense against pathogens or predators. |
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Keywords: | Protease inhibitor Mycocypin Coprinopsis cinerea Clitocybe nebularis Macrolepiota procera Expression |
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