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Pseudomonas putida

ABSTRACT: Development of multilocus sequence typing (MLST) for characterization of Pseudomonas putida

PROVIDER: PRJDB7626 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	DRR157750_1.fastq.gz	Fastqsanger.gz
	DRR157750_2.fastq.gz	Fastqsanger.gz
	DRR157751_1.fastq.gz	Fastqsanger.gz
	DRR157751_2.fastq.gz	Fastqsanger.gz
	DRR157752_1.fastq.gz	Fastqsanger.gz

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Project description:KaiC is the central cog of the circadian clock in Cyanobacteria. Close homologs of this protein are widespread among bacteria not known to have a circadian physiology. The function, interaction network, and mechanism of action of these KaiC homologs are still largely unknown. Here, we focus on KaiC homologs found in environmental Pseudomonas species. We characterize experimentally the only KaiC homolog present in Pseudomonas putida KT2440 and Pseudomonas protegens CHA0. Through phenotypic assays and transcriptomics, we show that KaiC is involved in osmotic and oxidative stress resistance in P. putida and in biofilm production in both P. putida and P. protegens.

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Project description:Pseudomonas putida KT2440 encodes 3 homologs of the LitR/CarH family (designated PplR1–PplR3; Pseudomonas putida light-induced transcription; regulator), which is an adenosyl B12-dependent light-sensitive MerR family transcriptional regulator. Transcriptome and individual transcriptional analysis revealed the existence of a number of photo-inducible genes including pplR1, phrB (encoding DNA photolyase), cfaA (cyclopropane synthase), folE (GTP cyclohydrolase I), cryB (cryptochrome-like protein), and multiple hypothetical genes. Transcriptional analysis based on a β-galactosidase reporter assay with single-, double-, and triple-knockout mutants of pplR1–pplR3 showed that deletion of pplR1–pplR3 completely abolished the light-inducible transcription in P. putida, which indicates that the transcription of light-inducible genes is under the ternary regulation of PplR proteins. DNase I footprint assay showed that PplR1 protein specifically binds to the promoter regions of light-inducible genes, suggesting a consensus PplR1-binding site, 5’-T(G/A)TACAn12TGTA(C/T)A-3’, predicted upon nucleotide sequence alignment. The disruption of cobalamin biosynthesis cluster did not affect the light-inducible transcription; however, disruption of ppSB1-LOV and ppSB2-LOV, a blue light photoreceptor genes, which are adjacent to pplR3 and pplR2, respectively, led to the complete loss of light-inducible transcription. Overall, the results suggest that 3 PplR and 2 PpSB-LOV regulate light-induced gene transcription in response to illumination. The high conservation of the pplR/ppSB-LOV cognate pair in Pseudomonas spp. suggests that the response and adaptation to light is similarly regulated in the group of non-phototrophic bacteria.

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