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RNA-seq reveals the RNA binding proteins, Hfq and RsmA, play various roles in virulence, antibiotic production and genomic flux in Serratia sp. ATCC 39006.

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Title: RNA-seq reveals the RNA binding proteins, Hfq and RsmA, play various roles in virulence, antibiotic production and genomic flux in Serratia sp. ATCC 39006.
Authors: Wilf NM, Reid AJ, Ramsay JP, Williamson NR, Croucher NJ, Gatto L, Hester SS, Goulding D, Barquist L, Lilley KS, Kingsley RA, Dougan G, Salmond GP; Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QW, UK. gpcs2@cam.ac.uk.
Source: BMC genomics [BMC Genomics] 2013 Nov 22; Vol. 14, pp. 822. Date of Electronic Publication: 2013 Nov 22.
Publication Type: Journal Article; Research Support, Non-U.S. Gov't
Language: English
Journal Info: Publisher: BioMed Central Country of Publication: England NLM ID: 100965258 Publication Model: Electronic Cited Medium: Internet ISSN: 1471-2164 (Electronic) Linking ISSN: 14712164 NLM ISO Abbreviation: BMC Genomics Subsets: MEDLINE
Imprint Name(s): Original Publication: London : BioMed Central, [2000-
MeSH Terms: Anti-Bacterial Agents/*biosynthesis , Bacterial Proteins/*genetics , Bacterial Proteins/*metabolism , Host Factor 1 Protein/*genetics , Host Factor 1 Protein/*metabolism , Serratia/*genetics , Serratia/*metabolism, Bacteriophages/genetics ; Bacteriophages/metabolism ; Electron Transport/genetics ; Flagella/genetics ; Flagella/metabolism ; Prodigiosin/metabolism ; RNA-Binding Proteins/genetics ; RNA-Binding Proteins/metabolism ; Serratia/pathogenicity ; Serratia/virology ; Virulence/genetics ; Virulence Factors/genetics ; Virulence Factors/metabolism ; Cluster Analysis ; Gene Expression Regulation, Bacterial ; Genome, Bacterial ; Mutation ; Operon ; Proteome ; Regulatory Sequences, Nucleic Acid ; Sequence Analysis, RNA ; Transcriptome
Abstract: Background: Serratia sp. ATCC 39006 (S39006) is a Gram-negative enterobacterium that is virulent in plant and animal models. It produces a red-pigmented trypyrrole secondary metabolite, prodigiosin (Pig), and a carbapenem antibiotic (Car), as well as the exoenzymes, pectate lyase and cellulase. Secondary metabolite production in this strain is controlled by a complex regulatory network involving quorum sensing (QS). Hfq and RsmA (two RNA binding proteins and major post-transcriptional regulators of gene expression) play opposing roles in the regulation of several key phenotypes within S39006. Prodigiosin and carbapenem production was abolished, and virulence attenuated, in an S39006 ∆hfq mutant, while the converse was observed in an S39006 rsmA transposon insertion mutant.
Results: In order to define the complete regulon of Hfq and RsmA, deep sequencing of cDNA libraries (RNA-seq) was used to analyse the whole transcriptome of S39006 ∆hfq and rsmA::Tn mutants. Moreover, we investigated global changes in the proteome using an LC-MS/MS approach. Analysis of differential gene expression showed that Hfq and RsmA directly or indirectly regulate (at the level of RNA) 4% and 19% of the genome, respectively, with some correlation between RNA and protein expression. Pathways affected include those involved in antibiotic regulation, virulence, flagella synthesis, and surfactant production. Although Hfq and RsmA are reported to activate flagellum production in E. coli and an adherent-invasive E. coli hfq mutant was shown to have no flagella by electron microscopy, we found that flagellar production was increased in the S39006 rsmA and hfq mutants. Additionally, deletion of rsmA resulted in greater genomic flux with increased activity of two mobile genetic elements. This was confirmed by qPCR and analysis of rsmA culture supernatant revealed the presence of prophage DNA and phage particles. Finally, expression of a hypothetical protein containing DUF364 increased prodigiosin production and was controlled by a putative 5' cis-acting regulatory RNA element.
Conclusion: Using a combination of transcriptomics and proteomics this study provides a systems-level understanding of Hfq and RsmA regulation and identifies similarities and differences in the regulons of two major regulators. Additionally our study indicates that RsmA regulates both core and variable genome regions and contributes to genome stability.
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Grant Information: BB/H013261/1 United Kingdom BB_ Biotechnology and Biological Sciences Research Council; United Kingdom WT_ Wellcome Trust
Substance Nomenclature: 0 (Anti-Bacterial Agents)
0 (Bacterial Proteins)
0 (Host Factor 1 Protein)
0 (Proteome)
0 (RNA-Binding Proteins)
0 (Virulence Factors)
OL369FU7CJ (Prodigiosin)
Entry Date(s): Date Created: 20131126 Date Completed: 20140911 Latest Revision: 20250529
Update Code: 20260130
PubMed Central ID: PMC4046660
DOI: 10.1186/1471-2164-14-822
PMID: 24267595
Database: MEDLINE
Description
ISSN:1471-2164
DOI:10.1186/1471-2164-14-822