Physiological and transcriptomic characterization of a <i>fliA</i> mutant of <i>Pseudomonas putida</i> KT2440

Rodríguez‐Herva, José J.; Duque, Estrella; Molina‐Henares, María Antonia; Navarro‐Avilés, Gloria; Dillewijn, Pieter van; Torre, J. de la; Molina‐Henares, Antonio J.; Campa, Ana M. Sánchez de la; Ran, F. Ann; Segura, Ana; Shingler, Victoria; Ramos, Juan L.

doi:10.1111/j.1758-2229.2009.00084.x

Cited by 29 publications

(50 citation statements)

References 42 publications

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“…Consistent with previously published results showing that transcription of bifA in a fliA mutant decreased approximately twofold in a microarray analysis (Rodríguez‐Herva et al., ), we observed twofold lower levels of bifA promoter transcription activity in fliA mutant compared with that of the wild type (Fig. B).…”

Section: Resultssupporting

confidence: 93%

“…This implied that FliA putatively functions as a c‐di‐GMP regulator. It is interesting to see that in a previous microarray analysis showed that the transcription of bifA in a P. putida fliA mutant decreased approximately twofold (Rodríguez‐Herva et al., ). Therefore, we hypothesized that the expression of BifA was putatively directly regulated by FilA and FliA possessed an ability to modulate the intracellular c‐di‐GMP level via regulating BifA expression in P. putida .…”

Section: Introductionmentioning

confidence: 97%

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Expression of the phosphodiesterase BifA facilitating swimming motility is partly controlled by FliA inPseudomonas putidaKT2440

et al. 2016

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Flagella‐mediated motility is an important capability of many bacteria to survive in nutrient‐depleted and harsh environments. Decreasing the intracellular cyclic di‐GMP (c‐di‐GMP) level by overexpression of phosphodiesterase BifA promotes flagellar‐mediated motility and induces planktonic lifestyle in Pseudomonas. The mechanism that regulates expression of bifA gene was poorly studied. Here we showed that expression of BifA was partly controlled by flagellar sigma factor FliA (σ28) in Pseudomonas putida KT2440. FliA deletion led to an approximately twofold decrease in transcription of bifA. 5′ race assay revealed two transcription start points in bifA promoter region, with the putative σ70 and σ28 promoter sequences upstream, respectively. Point mutation in σ28 promoter region reduced transcriptional activity of the promoter in wild‐type KT2440, but showed no influence on that in fliA deletion mutant. FliA overexpression decreased the intracellular c‐di‐GMP level in a BifA‐dependent way, suggesting that FliA was able to modulate the intracellular c‐di‐GMP level and BifA function was required for the modulation. Besides, FliA overexpression enhanced swimming ability of wild‐type strain, while made no difference to the bifA mutant. Our results suggest that FliA acts as a negative regulator to modulate the c‐di‐GMP level via controlling transcription of bifA to facilitate swimming motility.

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Section: Resultssupporting

confidence: 93%

Section: Introductionmentioning

confidence: 97%

Expression of the phosphodiesterase BifA facilitating swimming motility is partly controlled by FliA inPseudomonas putidaKT2440

et al. 2016

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“…1A). This, together with the recent finding from a transcriptomics analysis that aer2 is downregulated in P. putida KT2440 lacking FliA (σ 28 ) (Rodriguez‐Herva et al. , 2009), strongly suggested that aer2 is transcribed by FliA‐RNAP.…”

Section: Resultsmentioning

confidence: 67%

“…, 2009), strongly suggested that aer2 is transcribed by FliA‐RNAP. To verify this, we generated monocopy aer2 transcriptional reporter strains using wild‐type P. putida KT2440 and its FliA null counterpart, P. putida KT2440 fliA ::Km, in which the fliA gene is disrupted by a kanamycin resistance gene cassette (Rodriguez‐Herva et al. , 2009).…”

Section: Resultsmentioning

confidence: 99%

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The σ‐factor FliA, ppGpp and DksA coordinate transcriptional control of the aer2 gene of Pseudomonas putida

Osterberg

Skärfstad

Shingler

2010

Environmental Microbiology

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SummaryHere the s-factor requirement for transcription of three similar, but differentially regulated, aer genes of Pseudomonas putida KT2440 is investigated. Previous work has shown that the three Aer proteins, like chemoreceptors, colocalize to a single pole in a CheA-dependent manner. Lack of Aer2 -the most abundant of these three proteins -mediates defects in metabolism-dependent taxis and aerotaxis, while lack of Aer1 or Aer3 has no apparent phenotype. We show, using wild-type and mutant P. putida derivatives combined with P. putida reconstituted FliA-(s 28 ) and s 70-dependent in vitro transcription assays, that transcription of aer2 is coupled to motility through the flagella s-factor FliA, while s 70 is responsible for transcription of aer1 and aer3. By comparing activities of the wild-type and mutant forms of the aer2 promoter, we present evidence (i) that transcription from FliAdependent Paer2 is enhanced by changes towards the Escherichia coli consensus for FliA promoters rather than towards that of P. putida, (ii) that the nature of the AT-rich upstream region is important for both output and s 70 discrimination of this promoter, and (iii) that Paer2 output is directly stimulated by the bacterial alarmone ppGpp and its cofactor DksA.

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FleQ regulates both the type VI secretion system and flagella in Pseudomonas putida

Wang

et al. 2017

Biotech and App Biochem

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Based on the analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry, flagellin FliC and a key member of the type VI secretion system Hcp1 were found to be secreted from Pseudomonas putida KT2442 cells in the late exponential phase, but in the early exponential phase only FliC was secreted. Relevant genes fleQ and rpoN were then individually deleted in P. putida KT2442, and the expression and transcription of fliC and hcp1 in these mutant strains were analyzed. In both △fleQ and △rpoN mutant cells, Hcp1 but not FliC was secreted in the late exponential phase, and it was secreted even in the early exponential phase. Transcriptomic analysis of △fleQ cells shows that most of the genes relevant to the type VI secretion system, including hcp1, were upregulated; several genes involved in 3',5'-cyclic diguanylic acid (c-di-GMP) synthesis and degradation were also significantly regulated. To further study the regulation process of the type VI secretion system, the phosphodiesterase BifA was overexpressed to lower the intracellular c-di-GMP level. The results demonstrate that FleQ positively regulates flagella biosynthesis but negatively regulates the type VI secretion system in P. putida. This finding is important for understanding the function and regulation of the type VI secretion system.

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Physiological and transcriptomic characterization of a fliA mutant of Pseudomonas putida KT2440

Cited by 29 publications

References 42 publications

Expression of the phosphodiesterase BifA facilitating swimming motility is partly controlled by FliA inPseudomonas putidaKT2440

Expression of the phosphodiesterase BifA facilitating swimming motility is partly controlled by FliA inPseudomonas putidaKT2440

The σ‐factor FliA, ppGpp and DksA coordinate transcriptional control of the aer2 gene of Pseudomonas putida

FleQ regulates both the type VI secretion system and flagella in Pseudomonas putida

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