title: Rhodococcus erythropolis BG43 Genes Mediating Pseudomonas aeruginosa Quinolone Signal Degradation and Virulence Factor Attenuation.
abstract: Rhodococcus erythropolis BG43 is able to degrade the Pseudomonas aeruginosa quorum sensing signal molecules PQS (Pseudomonas quinolone signal) [2-heptyl-3-hydroxy-4(1H)-quinolone] and HHQ [2-heptyl-4(1H)-quinolone] to anthranilic acid. Based on the hypothesis that degradation of HHQ might involve hydroxylation to PQS followed by dioxygenolytic cleavage of the heterocyclic ring and hydrolysis of the resulting N-octanoylanthranilate, the genome was searched for corresponding candidate genes. Two gene clusters, aqdA1B1C1 and aqdA2B2C2, each predicted to code for a hydrolase, a flavin monooxygenase, and a dioxygenase related to 1H-3-hydroxy-4-oxoquinaldine 2,4-dioxygenase, were identified on circular plasmid pRLCBG43 of strain BG43. Transcription of all genes was upregulated by PQS, suggesting that both gene clusters code for alkylquinolone-specific catabolic enzymes. An aqdR gene encoding a putative transcriptional regulator, which was also inducible by PQS, is located adjacent to the aqdA2B2C2 cluster. Expression of aqdA2B2C2 in Escherichia coli conferred the ability to degrade HHQ and PQS to anthranilic acid; however, for E. coli transformed with aqdA1B1C1, only PQS degradation was observed. Purification of the recombinant AqdC1 protein verified that it catalyzes the cleavage of PQS to form N-octanoylanthranilic acid and carbon monoxide and revealed apparent Km and kcat values for PQS of ∼27 μM and 21 s(-1), respectively. Heterologous expression of the PQS dioxygenase gene aqdC1 or aqdC2 in P. aeruginosa PAO1 quenched the production of the virulence factors pyocyanin and rhamnolipid and reduced the synthesis of the siderophore pyoverdine. Thus, the toolbox of quorum-quenching enzymes is expanded by new PQS dioxygenases.
authors: Institute of Molecular Microbiology and Biotechnology, University of Münster, Münster, Germany.
authors: Christine
authors: C
authors: Müller
authors: Institute of Molecular Microbiology and Biotechnology, University of Münster, Münster, Germany.
authors: Franziska S
authors: FS
authors: Birmes
authors: Center for Biotechnology (CeBiTec), Bielefeld University, Bielefeld, Germany.
authors: Christian
authors: C
authors: Rückert
authors: Center for Biotechnology (CeBiTec), Bielefeld University, Bielefeld, Germany.
authors: Jörn
authors: J
authors: Kalinowski
authors: Institute of Molecular Microbiology and Biotechnology, University of Münster, Münster, Germany fetzner@uni-muenster.de.
authors: Susanne
authors: S
authors: Fetzner
mesh_terms: D015964:Gene Expression Regulation, Bacterial; D006017:Glycolipids / Q000235:genetics / Q000378:metabolism; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D010957:Plasmids; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000378:metabolism; D011710:Pyocyanine / Q000235:genetics / Q000378:metabolism; D015363:Quinolones / Q000378:metabolism; D053038:Quorum Sensing; D012240:Rhodococcus / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D037521:Virulence Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D006017:Glycolipids; D009842:Oligopeptides; D015363:Quinolones; D037521:Virulence Factors; C418382:rhamnolipid; C042453:pyoverdin; D011710:Pyocyanine