Testosterone-inducible Regulator Is a Kinase That Drives Steroid Sensing and Metabolism in Comamonas testosteroni

Göhler, André; Guan, Xiong; Paulsen, Simone; Trentmann, Gabriele; Maser, Edmund

doi:10.1074/jbc.m710166200

Cited by 42 publications

(28 citation statements)

References 45 publications

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“…Most of the genes in this cluster are involved in aromatization and following degradation of the A-ring in steroid degradation [14] (Figure 11). testosteroni [17,[67][68][69]. This transcriptional regulator is a LuxR family protein and possesses helix-turn-helix motif at the C-terminal to bind DNA.…”

Section: -6 Aromatization Of the A-ring Accompanied By The Cleavagementioning

confidence: 99%

Steroid degradation in Comamonas testosteroni

Horinouchi

Hayashi

Kudo

2012

The Journal of Steroid Biochemistry and Molecular Biology

114

153

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Steroid degradation by Comamonas testosteroni and Nocardia restrictus have been intensively studied for the purpose of obtaining materials for steroid drug synthesis. C. testosteroni degrades side chains and converts single/double bonds of certain steroid compounds to produce androsta-1,4-diene 3,17-dione or the derivative. Following 9-hydroxylation leads to aromatization of the A-ring accompanied by cleavage of the B-ring, and aromatized A-ring is hydroxylated at C-4 position, cleaved at ∆4 by meta-cleavage, and divided into 2-hydroxyhexa-2,4-dienoic acid (A-ring) and 9,17-dioxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid (B,C,D-ring) by hydrolysis.Reactions and the genes involved in the cleavage and the following degradation of the A-ring are similar to those for bacterial biphenyl degradation, and 9,17-dioxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid degradation is suggested to be mainly -oxidation. Genes involved in A-ring aromatization and degradation form a gene cluster, and the genes involved in -oxidation of 9,17-dioxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid also comprise a large cluster of more than 10 genes. The DNA region between these two main steroid degradation gene clusters contain 3-hydroxysteroid dehydrogenase gene, ∆5,3-ketosteroid isomerase gene, genes for inversion of an -oriented-hydroxyl group to a -oriented-hydroxyl group at C-12 position of cholic acid, and genes possibly involved in the degradation of a side chain at C-17 position of cholic acid, indicating this DNA region of more than 100kb to be a steroid degradation gene hot spot of C. testosteroni.

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Section: -6 Aromatization Of the A-ring Accompanied By The Cleavagementioning

confidence: 99%

Steroid degradation in Comamonas testosteroni

Horinouchi

Hayashi

Kudo

2012

The Journal of Steroid Biochemistry and Molecular Biology

114

153

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“…There is considerable interest in the mechanism of the induction of steroid-catabolic enzyme expression and the basis of their transcriptional regulation. In previous investigations, 3␣-HSD/CR from C. testosteroni ATCC 11996 was identified to be induced by steroids such as testosterone, and the expression of this enzyme was controlled by several regulators, including the repressors RepA and RepB and positive regulator TeiR (6,38,39).…”

Section: Discussionmentioning

confidence: 99%

“…RepB was demonstrated to bind to the mRNA of 3␣-HSD/CR and to interfere with 3␣-HSD/CR translation (39). The teiR gene, encoding a positive regulator of steroid-degrading enzymes, including 3␣-HSD/CR, was identified to mediate steroid sensing and signaling in C. testosteroni ATCC 11996 via a kinase mechanism (6).…”

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confidence: 99%

Identification and Characterization of the LysR-Type Transcriptional Regulator HsdR for Steroid-Inducible Expression of the 3α-Hydroxysteroid Dehydrogenase/Carbonyl Reductase Gene in Comamonas testosteroni

Gong¹,

Guan²,

Maser³

2012

Appl Environ Microbiol

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3␣-Hydroxysteroid dehydrogenase/carbonyl reductase (3␣-HSD/CR) fromIn the present study, a new LysR-type transcriptional factor, HsdR, for 3␣-HSD/CR expression in C. testosteroni has been identified. The hsdR gene is located 2.58 kb downstream from hsdA on the C. testosteroni ATCC 11996 chromosome with an orientation opposite that of hsdA. The hsdR gene was cloned and recombinant HsdR protein was produced, as was anti-HsdR polyclonal antibodies. While heterologous transformation systems revealed that HsdR activates the expression of the hsdA gene, electrophoresis mobility shift assays showed that HsdR specifically binds to the hsdA promoter region. Interestingly, the activity of HsdR is dependent on decreased repression by RepA. Furthermore, in vitro binding assays indicated that HsdR can come into contact with RNA polymerase. As expected, an hsdR knockout mutant expressed low levels of 3␣-HSD/CR compared to that of wild-type C. testosteroni after testosterone induction. In conclusion, HsdR is a positive transcription factor for the hsdA gene and promotes the induction of 3␣-HSD/CR expression in C. testosteroni.

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“…To further test and verify the steroid biodegradability by strain H5, 3 H labeled testosterone and estradiol were used in the present study. C. testosteroni ATCC1196 was employed as positive control to confirm that the procedure involved is competent in proving the steroid biodegradability by strain H5 [20][21][22][23][31][32][33][34][35][36][37][38][39][40].…”

Section: Steroid Biodegradability By Marine Strain H5mentioning

confidence: 99%

“…Biodegradations by microorganisms thereby play an extremely vital role to solve this urgent contaminant issue [14][15][16][17][18][19]. Built upon our long experience with the soil bacterium Comamonas testosteroni (C. testosteroni) which is able to catabolize a variety of steroids and PAH [20][21][22][23], we now seek to identify and characterize new marine bacterial strains that might be used for the bioremediation of steroids in seawater.…”

Section: Introductionmentioning

confidence: 99%

Steroid degradation and two steroid-inducible enzymes in the marine bacterium H5

Sang

Guan²,

Maser³

2011

Chemico-Biological Interactions

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a b s t r a c tNatural and synthetic steroid hormones excreted into the environment are potentially threatening the population dynamics of all kinds of animals and public health. We have previously isolated a steroid degrading bacterial strain (H5) from the Baltic Sea, at Kiel, Germany. 16S-rRNA analysis showed that bacterial strain H5 belongs to the genus Vibrio, family Vibrionaceae and class Gamma-Proteobacteria. Bacterial strain H5 can degrade steroids such as testosterone and estrogens, which was shown in this study by determining the 3 H labeled steroid retaining in the bacterial H5 culture medium at incubation times of 5 h and 20 h. Since 3␣-hydroxysteroid dehydrogenase/carbonyl reductase (3␣-HSD/CR) is a key enzyme in adaptive steroid degradation in Comamonas testosteroni (C. testosteroni), in previous investigations, a meta-genomic system with the 3␣-HSD/CR gene as a positive control was established. By this meta-genomic system, two estradiol inducible genes coding 3-ketosteroid-delta-1-dehydrogenase and carboxylesterase, respectively, which are involved in steroid degradation, were found in marine strain H5. In the present work, the 3-ketosteroid-delta-1-dehydrogenase and carboxylesterase genes were subcloned into plasmids pET38-12 and pET24-17, respectively. Overexpression in Escherichia coli (E. coli) strain BL21(DE3)pLysS cells resulted in corresponding proteins with an N-terminal His-tag sequence. After induction with isopropyl-␤-D-thiogalactoside, 3-ketosteroid-delta-1-dehydrogenase and carboxylesterase were purified in one step using nickel-chelate chromatography. After protein determination, 3-ketosteroid-delta-1-dehydrogenase (0.48 mg/ml) and carboxylesterase (1.28 mg/ml) were used to prepare antibodies to determine steroid binding specificity in future research. In summary, we have shown that the marine strain H5 could metabolize steroids; have isolated two estradiol inducible genes from strain H5 chromosomal DNA, and purified the corresponding proteins for further research. The exact characterization and systematic classification of the marine steroid degrading bacterial strain H5 is envisaged. The strain might be used for the bioremediation of steroid contaminations in seawater.

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Testosterone-inducible Regulator Is a Kinase That Drives Steroid Sensing and Metabolism in Comamonas testosteroni

Cited by 42 publications

References 45 publications

Steroid degradation in Comamonas testosteroni

Steroid degradation in Comamonas testosteroni

Identification and Characterization of the LysR-Type Transcriptional Regulator HsdR for Steroid-Inducible Expression of the 3α-Hydroxysteroid Dehydrogenase/Carbonyl Reductase Gene in Comamonas testosteroni

Steroid degradation and two steroid-inducible enzymes in the marine bacterium H5

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