title: A novel 17beta-hydroxysteroid dehydrogenase in the fungus Cochliobolus lunatus: new insights into the evolution of steroid-hormone signalling.
abstract: 17beta-Hydroxysteroid dehydrogenase (17beta-HSD) from the filamentous fungus Cochliobolus lunatus (17beta-HSDcl) catalyses the reduction of steroids and of several o- and p-quinones. After purification of the enzyme, its partial amino acid sequence was determined. A PCR fragment amplified with primers derived from peptide sequences was generated for screening the Coch. lunatus cDNA library. Three independent full-length cDNA clones were isolated and sequenced, revealing an 810-bp open reading frame encoding a 270-amino-acid protein. After expression in Escherichia coli and purification to homogeneity, the enzyme was found to be active towards androstenedione and menadione, and was able to form dimers of Mr 60000. The amino acid sequence of the novel 17beta-HSD demonstrated high homology with fungal carbonyl reductases, such as versicolorin reductase from Emericella nidulans (Aspergillus nidulans; VerA) and Asp. parasiticus (Ver1), polyhydroxynaphthalene reductase from Magnaporthe grisea, the product of the Brn1 gene from Coch. heterostrophus and a reductase from Colletotrichum lagenarium, which are all members of the short-chain dehydrogenase/reductase superfamily. 17beta-HSDcl is the first discovered fungal 17beta-hydroxysteroid dehydrogenase belonging to this family. The primary structure of this enzyme may therefore help to elucidate the evolutionary history of steroid dehydrogenases.
authors.affiliation: Institute of Biochemistry, Medical Faculty, Vrazov Trg 2, 1000 Ljubljana, Slovenia. Lanisnik@ibmi.mf.uni-lj.si
authors.forename: T
authors.initials: T
authors.lastname: Lanisnik Rizner
authors.forename: G
authors.initials: G
authors.lastname: Moeller
authors.forename: H H
authors.initials: HH
authors.lastname: Thole
authors.forename: M
authors.initials: M
authors.lastname: Zakelj-Mavric
authors.forename: J
authors.initials: J
authors.lastname: Adamski
mesh_terms: D015067:17-Hydroxysteroid Dehydrogenases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000735:Androstenedione / Q000378:metabolism; D001203:Ascomycota / Q000201:enzymology*; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006728:Hormones / Q000502:physiology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013256:Steroids / Q000502:physiology*; D014812:Vitamin K / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D006728:Hormones; D013256:Steroids; D014812:Vitamin K; D000735:Androstenedione; D015067:17-Hydroxysteroid Dehydrogenases