Expression of the phosphodiesterase BifA facilitating swimming motility is partly controlled by FliA in<i>Pseudomonas putida</i>KT2440

Xiao, Yujie; Liu, Huizhong; Nie, Hailing; Xie, Shuguang; Luo, Xuesong; Chen, Wenli; Huang, Qiaoyun

doi:10.1002/mbo3.402

Cited by 9 publications

(9 citation statements)

References 38 publications

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“…BifA degrades c‐di‐GMP and lower intracellular c‐di‐GMP levels. Expression of the BifA is partly controlled by the flagellar sigma factor FliA in P. putida KT2440 . FliA deletion led to a twofold decrease in the transcriptional level of bifA .…”

Section: Discussionmentioning

confidence: 99%

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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Section: Discussionmentioning

confidence: 99%

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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“…The bacterial secondary messenger cyclic diguanylate (c-di-GMP) regulates multiple important functions, including the transition from a planktonic lifestyle to a biofilm lifestyle and the biosynthesis of exopolysaccharides in the extracellular matrix of biofilms in many bacterial species (1–5). c-di-GMP is catalyzed by diguanylate cyclases (DGCs) and degraded by phosphodiesterases (PDEs).…”

Section: Introductionmentioning

confidence: 99%

“…A high intracellular c-di-GMP level drastically inhibits the biosynthesis of flagellin and bacterial motility but enhances the formation of biofilm. In contrast, a low c-di-GMP level strengthens bacterial motility and attenuates biofilm production in many bacteria (2, 5, 11–14). In P.…”

Section: Introductionmentioning

confidence: 99%

Pleiotropic Effects of c-di-GMP Content in Pseudomonas syringae

Wang

Cai

Shao

et al. 2019

Appl Environ Microbiol

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Although the ubiquitous bacterial secondary messenger cyclic diguanylate (c-di-GMP) has important cellular functions in a wide range of bacteria, its function in the model plant pathogen Pseudomonas syringae remains largely elusive. To this end, we overexpressed Escherichia coli diguanylate cyclase (YedQ) and phosphodiesterase (YhjH) in P. syringae, resulting in high and low in vivo levels of c-di-GMP, respectively. Via genome-wide RNA sequencing of these two strains, we found that c-di-GMP regulates (i) fliN, fliE, and flhA, which are associated with flagellar assembly; (ii) alg8 and alg44, which are related to the exopolysaccharide biosynthesis pathway; (iii) pvdE, pvdP, and pvsA, which are associated with the siderophore biosynthesis pathway; and (iv) sodA, which encodes a superoxide dismutase. In particular, we identified three promoters that are sensitive to elevated levels of c-di-GMP and inserted them into luciferase-based reporters that respond effectively to the c-di-GMP levels in P. syringae; these promoters could be useful in the measurement of in vivo levels of c-di-GMP in real time. Further phenotypic assays validated the RNA sequencing (RNA-seq) results and confirmed the effect on c-di-GMP-associated pathways, such as repressing the type III secretion system (T3SS) and motility while inducing biofilm production, siderophore production, and oxidative stress resistance. Taken together, these results demonstrate that c-di-GMP regulates the virulence and stress response in P. syringae, which suggests that tuning its level could be a new strategy to protect plants from attacks by this pathogen. IMPORTANCE The present work comprehensively analyzed the transcriptome and phenotypes that were regulated by c-di-GMP in P. syringae. Given that the majority of diguanylate cyclases and phosphodiesterases have not been characterized in P. syringae, this work provided a very useful database for the future study on regulatory mechanism (especially its relationship with T3SS) of c-di-GMP in P. syringae. In particular, we identified three promoters that were sensitive to elevated c-di-GMP levels and inserted them into luciferase-based reporters that effectively respond to intracellular levels of c-di-GMP in P. syringae, which could be used as an economic and efficient way to measure relative c-di-GMP levels in vivo in the future.

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“…This result could not be explained by the general model. Besides, our former study demonstrated that introducing multiple copies of PP_0914 ( bifA ) under the control of its own promoter into wild type KT2440 caused increased swimming motility and decreased biofilm formation (Xiao et al ., ). Here, in this study, we introduced multiple copies of PP_0914 under the control of a tac promoter into wild type KT2440 but found no obvious influence on biofilm formation (Fig.…”

Section: Resultsmentioning

confidence: 97%

“…Thus, we wonder why the native and inducible promoters of PP_0914 lead to different phenotype. Our former study indicated that the expression of PP_0914 was controlled by both fliA and rpoD simultaneously (Xiao et al ., ), but detailed regulation of PP_0914 was still unclear. Future studies on the regulation of PP_0914 expression will help to answer this question.…”

Section: Resultsmentioning

confidence: 99%

Phenotypic–genotypic analysis of GGDEF/EAL/HD‐GYP domain‐encoding genes in Pseudomonas putida

Nie

Xiao

et al. 2019

Environ Microbiol Rep

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Cyclic diguanylate (c-di-GMP) is a broadly conserved bacterial signalling molecule that modulates diverse cellular processes, such as biofilm formation, colony morphology and swimming motility. The intracellular level of c-di-GMP is controlled by diguanylate cyclases (DGCs) with GGDEF domain and phosphodiesterases (PDEs) with either EAL or HD-GYP domain. Pseudomonas putida KT2440 has a large group of genes on its genome encoding proteins with GGDEF/EAL/HD-GYP domains. However, phenotypic-genotypic correlation and c-di-GMP metabolism of these genes were largely unknown. Herein, by systematically constructing deletion mutants/overexpression strains of the 42 predicted c-di-GMP metabolism-related genes and analysing the phenotypes, we preliminarily revealed the role of each gene in biofilm formation, colony morphology and swimming motility. Subsequent results from protein sequence alignments and cellular c-di-GMP assessment indicated that 25 out of the 42 genes were likely to encode DGCs, nine genes were predicted to encode PDEs, four genes encoded bifunctional enzymes and the other four genes encoded enzymatically inactive proteins. This study offers a basic understanding of the roles of these 42 genes and can serve as a toolkit for investigators to further elucidate the functions of these GGDEF and EAL/HD-GYP domain-containing proteins.

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

Cited by 9 publications

References 38 publications

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

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

Pleiotropic Effects of c-di-GMP Content in Pseudomonas syringae

Phenotypic–genotypic analysis of GGDEF/EAL/HD‐GYP domain‐encoding genes in Pseudomonas putida

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