Testosterone-inducible regulator is a kinase that drives steroid sensing and metabolism in Comamonas testosteroni.
The mechanism of gene regulation by steroids in bacteria is still a mystery. We use steroid-inducible 3alpha-hydroxysteroid dehydrogenase/carbonyl reductase (3alpha-HSD/CR) as a reporter system to study steroid signaling in Comamonas testosteroni. In previous investigations we cloned and characterized the 3alpha-HSD/CR-encoding gene, hsdA. In addition, we identified two negative regulator genes (repA and repB) in the vicinity of hsdA, the protein products which repress hsdA expression on the level of transcription and translation, respectively. Recently, a positive regulator of hsdA expression, TeiR (testosterone-inducible regulator), was found by transposon mutagenesis, but the mode of its action remained obscure. In the present work we produced a TeiR-green fluorescent fusion protein and showed that TeiR is a membrane protein with asymmetrical localization at one of the cell poles of C. testosteroni. Knock-out mutants of the teiR gene revealed that TeiR provides swimming and twitching motility of C. testosteroni to the steroid substrate source. TeiR also mediated an induced expression of 3alpha-HSD/CR which was paralleled by an enhanced catabolism of testosterone. We also found that TeiR responds to a variety of different steroids other than testosterone. Biochemical analysis with several deletion mutants of the teiR gene revealed TeiR to consist of three different functional domains, an N-terminal domain important for membrane association, a central steroid binding site, and a C-terminal part mediating TeiR function. Finally, we could demonstrate that TeiR works as a kinase in the steroid signaling chain in C. testosteroni. Overall, we provide evidence that TeiR mediates steroid sensing and metabolism in C. testosteroni via its steroid binding and kinase activity.
Göhler
André
A
Institute of Toxicology and Pharmacology for Natural Scientists, University Medical School Schleswig-Holstein, Campus Kiel, Brunswiker Strasse 10, 24105 Kiel, Germany.
Xiong
Guangming
G
Paulsen
Simone
S
Trentmann
Gabriele
G
Maser
Edmund
E
D001426:Bacterial Proteins / Q000378:metabolism / Q000502:physiology*; D003001:Cloning, Molecular; D020604:Comamonas testosteroni / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D008565:Membrane Proteins / Q000378:metabolism / Q000502:physiology*; D008954:Models, Biological; D008957:Models, Genetic; D009154:Mutation; D010770:Phosphotransferases / Q000378:metabolism / Q000502:physiology*; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D015398:Signal Transduction; D013256:Steroids / Q000737:chemistry / Q000378:metabolism*; D013739:Testosterone / Q000494:pharmacology*
D001426:Bacterial Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; D013256:Steroids; D013739:Testosterone; D010770:Phosphotransferases; C530261:TeiR protein, Comamonas testosteroni