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An XRE-superfamily transcriptional regulator XrpA controls low pH tolerance of Serratia marcescens

ABSTRACT: An XRE-superfamily transcriptional regulator XrpA controls low pH tolerance of Serratia marcescens

PROVIDER: PRJNA564772 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10093214_1.fastq.gz	Fastqsanger.gz
	SRR10093214_2.fastq.gz	Fastqsanger.gz
	SRR10093215_1.fastq.gz	Fastqsanger.gz
	SRR10093215_2.fastq.gz	Fastqsanger.gz

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An XRE-superfamily transcriptional regulator XrpA controls low pH tolerance of Serratia marcescens

Project description:The acid tolerance of industrial strains is a significant challenge in the fermentation process. The bacterium Serratia marcescens is part of the Enterobacteriaceae family of eubacteria, which is a potential industrial microorganism. However, the molecular mechanism behind S. marcescens acid resistance is not properly understood. In this study, we screened for novel regulators that respond to acidic conditions by a Tn5G transposon insertion mutagenesis of S. marcescens and found mutations in a gene encoding for the HTH_XRE super-family regulatory protein member, here named xrpA. Using transcriptomics and further experiments, we showed that the xrpA disruption conferred pleiotropic phenotype changes, including highly decreased biomass, altered cell shape, H+-ATPase activity, and deficiency of cell membrane permeability and integrity, compared with those of the parent (JNB5-1) strain at low pH. These data revealed that the molecular mechanism by which xrpA affects acid resistance of S. marcescens is through positive regulation of cell membrane permeability, integrity, and H+-ATPase activity to maintain intracellular homeostasis at low pH. Finally, we constructed an acidic resistant strain JNB5-1/pSX1314 by overexpression of xrpA in S. marcescens and found that its ability to produce prodigiosin by fermentation increased by 21.74% compared with that of parent strain at pH4.0. These results indicated that xrpA regulates tolerance to low pH by transcriptional regulation of acid stress response genes to maintain cell membrane function in S. marcescens.

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Project description:In order to identify changes in the global mRNA transcriptome caused by deletion of the RNA-binding protein Hfq in Serratia marcescens, total mRNA was isolated from wild type Serratia marcescens Db10 and an otherwise isogenic strain carrying an in-frame deletion of the hfq gene (SMDB11_4482) and analysed by RNAseq. Four independent biological replicates were sequenced for each strain using the Illumina HiSeq platform. The data was used to identify the nature and extent of changes in transcript level between the two strains and to inform on the role of Hfq in virulence of Serratia marcescens, an opportunist bacterial pathogen.

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Global identification of Hfq-bound RNA and transcriptional start site mapping in Serratia marcescens.

Project description:In order to identify mRNA and sRNAs associated with the RNA-binding protein Hfq in Serratia marcescens strain Db10, Hfq-bound RNA was immunoprecipitated from a strain encoding an Hfq-3FLAG fusion protein at the normal location and sequenced, in parallel with the wild type strain (no fusion) as negative control. Additionally global transcriptional start site mapping was performed on total RNA, with or without TEX treatment, isolated from wild type Serratia marcescens. The data was used to identify regions of mRNA and sRNAs associated with Hfq in this organism. Associated work in Serratia marcescens Db10, an opportunistic bacterial pathogen, has shown that Hfq is essential for virulence in several models and exerts a wide-ranging impact on the transcriptome and, particularly, genes encoding virulence factors.

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Project description:We report the application of transcriptome sequencing technology for high-throughput profiling of Serratia marcescens for producing prodigiosin. By obtaining over 163 million bases of sequence from Serratia marcescens genome DNA, we generated transcriptome -state maps of Serratia marcescens 12h cells, 24h cells, and 36h cells at 30C and 37C,respectively. We explored the mechanism of S. marcescens response temperature regulation at the transcription level through transcriptome sequencing technology. We found that the pig gene cluster at low temperature would favor at the transcriptional level, however, higher temperature resulting in instability and loss of enzyme activity. Numerous amino acid metabolic pathways involved in prodigiosin biosynthesis in S. marcescens responded to temperature changes, and metabolic fluxes were directed towards prodigiosin biosynthesis. At the same time, quorum sensing, two-component regulatory system and sRNA were stimulated by temperature to regulate PG biosynthesis and involve strain virulence and exclusive genes. Moreover, inhibition factors was the one reason for S. marcescens incapable synthesis of prodigiosin at 37C. This study laid a good foundation for understanding the biological functions of prodigiosin, improving the temperature tolerance of industrial strains, and excavating temperature-sensitive regulatory elements.

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