The Anti-activator QslA Negatively Regulates Phenazine-1-Carboxylic Acid Biosynthesis by Interacting With the Quorum Sensing Regulator MvfR in the Rhizobacterium Pseudomonas aeruginosa Strain PA1201

Fang, Yun-Ling; Chen, Bin; Zhou, Lian; Jin, Zi-Jing; Sun, Shuang; He, Yanan

doi:10.3389/fmicb.2018.01584

Cited by 6 publications

(5 citation statements)

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“…MvfR is one of the most important QS regulators in P. aeruginosa (Wade et al, 2005). The PQS signal receptor MvfR regulates 2-heptyl-4-quinolone (HHQ) and PQS biosynthesis by binding the promoter of pqsABCDE (Wade et al, 2005;Fang et al, 2018). MvfR is essential for PCA biosynthesis in strain P. aeruginosa PA1201 (Sun et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Phenazine and quinolone are main compounds produced by Pseudomonas aeruginosa N1 during inhibitory interaction with Rhizopus microsporus

2023

ANRES

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Importance of the work: Pseudomonas aeruginosa N1 inhibits Rhizopus microsporus growth by the production of quinolone, phenazine and rhamnolipid. Objectives: To evaluate the compounds extracted from P. aeruginosa N1 monoculture and from co-culture between R. microsporus and P. aeruginosa N1 and to observe the degree of damage caused to R. microsporus by its inhibitory interaction with P. aeruginosa N1. Materials & Methods:The inhibitory interaction of P. aeruginosa N1 was studied based on the metabolite released during R. microsporus inhibition. The substance released during the inhibitory interaction between R. microsporus and P. aeruginosa N1 was compared using monoculture and co-culture methods, with further analysis using liquid chromatography mass spectrophotometry (LC-MS). Results: Based on the LC-MS analysis, quinolone, phenazine and rhamnolipid compounds were the main substances produced during the interaction between species. There was a slight difference between the detected compounds produced in monocultures and those produced in co-culture. Specifically, 2-nonyl-4-hydroxyquinoline N-oxide and monorhamnolipid compounds were detected only in the P. aeruginosa N1 monocultures. On the other hand, the compounds 2-heptyl-4-quinolone, 1-hydroxy-2-[(2E)-2-nonen-1-yl]-4 (1H)-quinolinone and 2-nonyl-3-hydroxy-4-quinolone, were detected only in co-culture. Main finding: Quinolone, phenazine and rhamnolipid were the main compounds produced during the inhibitory interaction between P. aeruginosa N1 and R. microsporus. Furthermore, these compounds played an important role in improving antagonism against R. microsporus. However, this may regulate differently in each strain.

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Section: Discussionmentioning

confidence: 99%

Phenazine and quinolone are main compounds produced by Pseudomonas aeruginosa N1 during inhibitory interaction with Rhizopus microsporus

2023

ANRES

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“…Phenazines are one of the most important metabolites produced by P. aeruginosa and play diverse roles in cellular growth, development, and virulence. The regulation of the phenazine PYO in P. aeruginosa has been thoroughly studied in the clinically isolated PAO1 strain [ 19 , 20 , 28 , 29 , [34] , [35] , [36] , [37] , [38] ] and both MvaT and MvaU have been demonstrated to negatively regulate PYO production in PAO1 [ 30 ]. Both mvaT and mvaU are present in PA1201.…”

Section: Discussionmentioning

confidence: 99%

“…Cells from fresh overnight cultures of P. aeruginosa PA1201 and its derivative strains were inoculated into 50 mL of PPM in a 250 mL Erlenmeyer flask at a final concentration equivalent to an optical density at 600 nm (OD 600 ) of 0.025, followed by incubation at 28 °C with shaking at 200 rpm. PCA was extracted with chloroform (pH 4.0) and quantified with high performance liquid chromatography (HPLC), as previously described [ 25 , 29 ]. HPLC with a UV–Vis detector was used for qualitative and quantitative analysis of PCA in the reverse phase on an Agilent Technologies 1260 Infinity instrument.…”

Section: Methodsmentioning

confidence: 99%

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H-NS family protein MvaU downregulates phenazine-1-carboxylic acid (PCA) biosynthesis via binding to an AT-rich region within the promoter of the phz2 gene cluster in the rhizobacterium Pseudomonas strain PA1201

Fang

Cui

Zhou

et al. 2021

Synthetic and Systems Biotechnology

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Histone-like nucleoid-structuring (H-NS) proteins are key regulators in gene expression silencing and in nucleoid compaction. The H-NS family member proteins MvaU in Pseudomonas aeruginosa are thought to bind the same AT-rich regions of chromosomes and function to coordinate the control of a common set of genes. Here, we explored the molecular mechanism by which MvaU controls PCA biosynthesis in P. aeruginosa PA1201. We present evidence suggesting that MvaU is self-regulated. Deletion of mvaU significantly increased PCA production, and PCA production sharply decreased when mvaU was over-expressed. MvaU transcriptionally repressed phz2 cluster expression and consequently reduced PCA biosynthesis. β-galactosidase assays confirmed that base pairing near the −35 box is required when MvaU regulates PCA production in PA1201. Electrophoretic mobility shift assays (EMSA) and additional point mutation analysis demonstrated that MvaU directly bound to an AT-rich motif within the promoter of the phz2 cluster. Chromatin immunoprecipitation (ChIP) analysis also indicated that MvaU directly bound to the P5 region of the phz2 cluster promoter. MvaU repression of PCA biosynthesis was independent of QscR and OxyR in PA1201 and neither PCA or H 2 O 2 were the environmental signals that induced mvaU expression. These findings detail a new MvaU-dependent regulatory pathway of PCA biosynthesis in PA1201 and provide a foundation to increase PCA fermentation titer by genetic engineering.

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“…Two duplicated gene clusters, phzA 1 B 1 C 1 D 1 E 1 F 1 G 1 ( phz1 ) and phzA 2 B 2 C 2 D 2 E 2 F 2 G 2 ( phz2 ), are responsible for PCA biosynthesis in both Pseudomonas strains . Over the last 10 years, PCA biosynthesis in M18 or PA1201 has been shown to be regulated by an untranslated region (UTR) in the phz1 promoter region, by three quorum sensing (QS) systems (LasI/LasR, RhlI/RhlR, and PQS/PQSR), and by the transcription factors RsaL, QslA, and OxyR. − On the basis of these findings, at least five rounds of genetic engineering have been conducted in both PA1201 and M18, and the PCA fermentation titer in the engineered strain PA-IV was reported to reach 9.8 g/L . PCA was commercially rebranded as Shenqinmycin in 2005.…”

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confidence: 99%

Identification of a Strong Quorum Sensing- and Thermo-Regulated Promoter for the Biosynthesis of a New Metabolite Pesticide Phenazine-1-carboxamide in Pseudomonas strain PA1201

et al. 2020

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Phenazine-1-carboxamide (PCN) produced by multifarious Pseudomonas strains represents a promising candidate as a new metabolite pesticide due to its broad-spectrum antifungal activity and capacity to induce systemic resistance in plants. The rice rhizosphere Pseudomonas strain PA1201 contains two reiterated gene clusters, phz1 and phz2, for phenazine-1-carboxylic acid (PCA) biosynthesis; PCA is further converted into PCN by this strain using a functional phzH-encoding glutamine aminotransferase. However, PCN levels in PA1201 constitute approximately one-fifth of PCA levels and the optimal temperature for PCN synthesis is 28 °C. In this study, the phzH open reading frame (ORF) and promoter region were investigated and reannotated. phzH promoter P phzH was found to be a weak promoter, and PhzH levels were not sufficient to convert all of the native PCA into PCN. Following RNA Seq and promoter-lacZ fusion analyses, a strong quorum sensing (QS)-and thermo-regulated promoter P rhlI was identified and characterized. The activity of P phzH is approximately 1% of P rhlI in PA1201. After three rounds of promoter editing and swapping by P rhlI , a new PCNoverproducing strain UP46 was generated. The optimal fermentation temperature for PCN biosynthesis in UP46 was increased from 28 to 37 °C and the PCN fermentation titer increased 179.5-fold, reaching 14.1 g/L, the highest ever reported.

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The Anti-activator QslA Negatively Regulates Phenazine-1-Carboxylic Acid Biosynthesis by Interacting With the Quorum Sensing Regulator MvfR in the Rhizobacterium Pseudomonas aeruginosa Strain PA1201

Cited by 6 publications

References 46 publications

Phenazine and quinolone are main compounds produced by Pseudomonas aeruginosa N1 during inhibitory interaction with Rhizopus microsporus

Phenazine and quinolone are main compounds produced by Pseudomonas aeruginosa N1 during inhibitory interaction with Rhizopus microsporus

H-NS family protein MvaU downregulates phenazine-1-carboxylic acid (PCA) biosynthesis via binding to an AT-rich region within the promoter of the phz2 gene cluster in the rhizobacterium Pseudomonas strain PA1201

Identification of a Strong Quorum Sensing- and Thermo-Regulated Promoter for the Biosynthesis of a New Metabolite Pesticide Phenazine-1-carboxamide in Pseudomonas strain PA1201

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