title: Gentisate and 3-oxoadipate pathways in the yeast Candida parapsilosis: identification and functional analysis of the genes coding for 3-hydroxybenzoate 6-hydroxylase and 4-hydroxybenzoate 1-hydroxylase.
abstract: The pathogenic yeast Candida parapsilosis degrades various hydroxy derivatives of benzenes and benzoates by the gentisate and 3-oxoadipate pathways. We identified the genes MNX1, MNX2, MNX3, GDX1, HDX1 and FPH1 that code for enzymes involved in these pathways in the complete genome sequence of C. parapsilosis. Next, we demonstrated that MNX1, MNX2, MNX3 and GDX1 are inducible and transcriptionally controlled by hydroxyaromatic substrates present in cultivation media. Our results indicate that MNX1 and MNX2 code for flavoprotein monooxygenases catalysing the first steps in the 3-oxoadipate and gentisate pathways, respectively (i.e. 4-hydroxybenzoate 1-hydroxylase and 3-hydroxybenzoate 6-hydroxylase). Moreover, we found that the two pathways differ by their intracellular localization. The enzymes of the 3-oxoadipate pathway, Mnx1p and Mnx3p, localize predominantly in the cytosol. In contrast, intracellular localization of the components of the gentisate pathway, Mnx2p and Gdx1p, depends on the substrate in the cultivation medium. In cells growing on glucose these proteins localize in the cytosol, whereas in media containing hydroxyaromatic compounds they associate with mitochondria. Finally, we showed that the overexpression of MNX1 or MNX2 increases the tolerance of C. parapsilosis cells to the antifungal drug terbinafine.
authors.affiliation: Department of Genetics, Comenius University in Bratislava, Faculty of Natural Sciences, Mlynska dolina B-1, 842 15 Bratislava, Slovak Republic.|Department of Biochemistry, Comenius University in Bratislava, Faculty of Natural Sciences, Mlynska dolina CH-1, 842 15 Bratislava, Slovak Republic.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Holesova
authors.affiliation: Department of Biochemistry, Comenius University in Bratislava, Faculty of Natural Sciences, Mlynska dolina CH-1, 842 15 Bratislava, Slovak Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Jakubkova
authors.affiliation: Department of Genetics, Comenius University in Bratislava, Faculty of Natural Sciences, Mlynska dolina B-1, 842 15 Bratislava, Slovak Republic.
authors.forename: Ivana
authors.initials: I
authors.lastname: Zavadiakova
authors.affiliation: Department of Biochemistry, Comenius University in Bratislava, Faculty of Natural Sciences, Mlynska dolina CH-1, 842 15 Bratislava, Slovak Republic.
authors.forename: Igor
authors.initials: I
authors.lastname: Zeman
authors.affiliation: Department of Genetics, Comenius University in Bratislava, Faculty of Natural Sciences, Mlynska dolina B-1, 842 15 Bratislava, Slovak Republic.
authors.forename: Lubomir
authors.initials: L
authors.lastname: Tomaska
authors.affiliation: Department of Biochemistry, Comenius University in Bratislava, Faculty of Natural Sciences, Mlynska dolina CH-1, 842 15 Bratislava, Slovak Republic.
authors.forename: Jozef
authors.initials: J
authors.lastname: Nosek
mesh_terms: D000272:Adipates / Q000378:metabolism*; D000935:Antifungal Agents / Q000494:pharmacology; D002175:Candida / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D025141:Drug Resistance, Fungal; D016681:Genome, Fungal; D005841:Gentisates / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D062385:Hydroxybenzoates / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D009281:Naphthalenes / Q000494:pharmacology; D010101:Oxygen Consumption; D019800:Phenol / Q000378:metabolism; D000077291:Terbinafine
chemical_list: D000272:Adipates; D000935:Antifungal Agents; D005841:Gentisates; D062385:Hydroxybenzoates; D009281:Naphthalenes; C027316:3-oxoadipic acid; D019800:Phenol; D006899:Mixed Function Oxygenases; C108841:4-hydroxybenzoate 1-hydroxylase (decarboxylating); C023315:3-hydroxybenzoate-6-hydroxylase; D000077291:Terbinafine; D005947:Glucose