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  • Article  (20)
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  • 1
    In: Molecular Microbiology, April 2012, Vol.84(1), pp.1-5
    Description: The transcription factor CsgD governing the production of curli fimbriae and cellulose is a key player in the complex regulatory circuit that decides whether form biofilms. The gene itself is tightly controlled at the level of transcription by a large array of DNA‐binding proteins, but what happens after transcription is less understood. In this issue of , Jørgensen (2012), Mika (2012) and Thomason (2012) report on small RNAs (McaS, RprA and GcvB) that together with the RNA‐chaperone Hfq regulate the mRNAs of and other biofilm genes, and illustrate the burgeoning concept that the 5′ region of bacterial mRNA serves as a hub for sRNA‐mediated signal integration at the post‐transcriptional level.
    Keywords: Transcription (Genetics) ; Proteins ; Messenger Rna ; Genes ; Cellulose;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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  • 2
    In: Molecular Microbiology, December 2010, Vol.78(6), pp.1327-1331
    Description: Although most bacterial small RNAs act to repress target mRNAs, some also activate messengers. The predominant mode of activation has been seen in ‘anti‐antisense’ regulation whereby a small RNA prevents the formation of an inhibitory 5′ mRNA structure that otherwise impairs translational initiation and protein synthesis. The translational activation might also stabilize the target yet this was considered a secondary effect in the examples known thus far. Two recent papers in investigate post‐transcriptional activation of collagenase mRNA by VR‐RNA, and streptokinase mRNA by FasX RNA, to suggest that small RNAs exert positive regulation of virulence genes primarily at the level of mRNA stabilization.
    Keywords: Protein Synthesis ; Messenger Rna;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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  • 3
    In: Molecular Microbiology, September 2009, Vol.73(5), pp.737-741
    Description: Small regulatory RNAs (sRNAs) are well known to command bacterial protein synthesis by modulating the translation and decay of target mRNAs. Most sRNAs are specifically regulated by a cognate transcription factor under certain growth or stress conditions. Investigations of the conserved Hfq‐dependent MicM sRNA in (article by Poul Valentin‐Hansen and colleagues in this issue of ) and in have unravelled a novel type of gene regulation in which the chitobiose operon mRNA acts as an RNA trap to degrade the constitutively expressed MicM sRNA, thereby alleviating MicM‐mediated repression of the synthesis of the YbfM porin that is required for chitosugar uptake. The results suggest that ‘target’ mRNAs might be both prey and also predators of sRNAs.
    Keywords: Protein Synthesis ; Messenger Rna;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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  • 4
    In: Molecular Microbiology, January 2009, Vol.71(1), pp.1-11
    Description: species are enterobacterial pathogens that have been exceptionally well investigated with respect to virulence mechanisms, microbial pathogenesis, genome evolution and many fundamental pathways of gene expression and metabolism. While these studies have traditionally focused on protein functions, has also become a model organism for RNA‐mediated regulation. The present review is dedicated to the non‐coding RNA world of : it covers small RNAs (sRNAs) that act as post‐transcriptional regulators of gene expression, novel Salmonella ‐regulatory RNA elements that sense metabolite and metal ion concentrations (or temperature), and globally acting RNA‐binding proteins such as CsrA or Hfq (inactivation of which cause drastic phenotypes and virulence defects). Owing to mosaic genome structure, some of the sRNAs are widely conserved in bacteria whereas others are very specific to species. Intriguingly, sRNAs of either type (CsrB/C, InvR, SgrS) facilitate cross‐talk between the core genome and its laterally acquired virulence regions. Work in also identified physiological functions (and mechanisms thereof) of RNA that had remained unknown in , and pioneered the use of high‐throughput sequencing technology to identify the sRNA and mRNA targets of bacterial RNA‐binding proteins.
    Keywords: Metabolites ; Proteins ; Messenger Rna ; Salmonella ; Gene Expression;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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  • 5
    In: Molecular Microbiology, September 2011, Vol.81(5), pp.1144-1165
    Description: GcvB is one of the most highly conserved Hfq‐associated small RNAs in Gram‐negative bacteria and was previously reported to repress several ABC transporters for amino acids. To determine the full extent of GcvB‐mediated regulation in , we combined a genome‐wide experimental approach with biocomputational target prediction. Comparative pulse expression of wild‐type versus mutant sRNA variants revealed that GcvB governs a large post‐transcriptional regulon, impacting ∼1% of all genes via its conserved G/U‐rich domain R1. Complementary predictions of C/A‐rich binding sites in mRNAs and reporter fusion experiments increased the number of validated GcvB targets to more than 20, and doubled the number of regulated amino acid transporters. Unlike the previously described targeting via the single R1 domain, GcvB represses the glycine transporter CycA by exceptionally redundant base‐pairing. This novel ability of GcvB is focused upon the one target that could feedback‐regulate the glycine‐responsive synthesis of GcvB. Several newly discovered mRNA targets involved in amino acid metabolism, including the global regulator Lrp, question the previous assumption that GcvB simply acts to limit unnecessary amino acid uptake. Rather, GcvB rewires primary transcriptional control circuits and seems to act as a distinct regulatory node in amino acid metabolism.
    Keywords: Glycine -- Physiological Aspects ; Genetic Research -- Physiological Aspects ; Genomics -- Physiological Aspects ; Messenger Rna -- Physiological Aspects;
    ISSN: 0950-382X
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  • 6
    In: Molecular Microbiology, October 2009, Vol.74(2), pp.261-269
    Description: A recent meeting on ‘Regulatory RNAs in prokaryotes’ reflected the growing interest in this research topic. Almost 200 scientists met to discuss the identification, structure, function and mechanistic details of regulatory RNAs in bacteria and archaea. The topics included small regulatory RNAs, riboswitches, RNA thermosensors and CRISPR (lustered egularly nterspaced hort alindromic epeats) elements.
    Keywords: Rna;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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  • 7
    In: Molecular Microbiology, December 2009, Vol.74(6), pp.1497-1512
    Description: Exposure to oxygen and light generates photooxidative stress by the bacteriochlorophyll mediated formation of singlet oxygen (O) in . Our study reports the genome‐wide search for small RNAs (sRNAs) involved in the regulatory response to O. By using 454 pyrosequencing and Northern blot analysis, we identified 20 sRNAs from aerobic cultures or following treatment with O or superoxide (O). One sRNA was specifically induced by O and its expression depends on the extracytoplasmic function sigma factor RpoE. Two sRNAs induced by O and O were cotranscribed with upstream genes preceded by promoters with target sequences for the alternative sigma factors RpoH and RpoH. The most abundant sRNA was processed in the presence of O but not by O. From this and a second sRNA a conserved 3′‐segment accumulated from a larger precursor. Absence of the RNA chaperone Hfq changed the half‐lives, abundance and processing of O‐affected sRNAs. Orthologues of three sRNA genes are present in different alpha‐proteobacteria, but the majority was unique to or species. Our discovery that abundant sRNAs are affected by O exposure extends the knowledge on the role of sRNAs and Hfq in the regulatory response to oxidative stress.
    Keywords: Active Oxygen -- Analysis ; Genomics -- Analysis ; Molecular Chaperones -- Analysis ; Rna -- Analysis ; Superoxides -- Analysis;
    ISSN: 0950-382X
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  • 8
    In: Molecular Microbiology, October 2009, Vol.74(1), pp.139-158
    Description: The small RNA, ArcZ (previously RyhA/SraH), was discovered in several genome‐wide screens in and . Its high degree of genomic conservation, its frequent recovery by shotgun sequencing, and its association with the RNA chaperone, Hfq, identified ArcZ as an abundant enterobacterial ‘core’ small RNA, yet its function remained unknown. Here, we report that ArcZ acts as a post‐transcriptional regulator in , repressing the mRNAs of the widely distributed (serine uptake) and (oxidative stress) genes, and of STM3216, a horizontally acquired methyl‐accepting chemotaxis protein (MCP). Both and STM3216 are regulated by sequestration of the ribosome binding site. In contrast, the mRNA is targeted in the coding sequence (CDS), arguing that CDS targeting is more common than appreciated. Transcriptomic analysis of an deletion strain further argued for the existence of a distinct set of loci specifically regulated by ArcZ. In contrast, increased expression of the sRNA altered the steady‐state levels of 〉 16% (〉 750) of all mRNAs, and rendered the bacteria non‐motile. Deep sequencing detected a dramatically changed profile of Hfq‐bound sRNAs and mRNAs, suggesting that the unprecedented pleiotropic effects by a single sRNA might in part be caused by altered post‐transcriptional regulation.
    Keywords: Biology;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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  • 9
    In: Molecular Microbiology, May 2008, Vol.68(4), pp.890-906
    Description: Post‐transcriptional repression of porin synthesis has emerged as a major function of Hfq‐dependent, small non‐coding RNAs (sRNAs). Many enterobacteria express OmpX‐like porins, a family of outer membrane proteins whose physiological roles and structural properties have been studied intensively. While regulatory sRNAs have been identified for most major and many minor porins of and , a post‐transcriptional regulator of OmpX levels has never been found. Here, we have taken a ‘reverse target search’ approach by systematic inactivation of sRNA genes, and screening 35 sRNA deletion strains for effects on OmpX synthesis. We have identified the Hfq‐dependent CyaR (formerly RyeE) sRNA as an repressor. Global transcriptomic profiling following induction of CyaR expression suggests that mRNA is the primary target of this sRNA under standard growth conditions. The results of phylogenetic and mutational analyses suggest that a conserved RNA hairpin of CyaR, featuring a C‐rich apical loop, acts to sequester the Shine–Dalgarno sequence of mRNA and to inhibit translational initiation. We have also discovered that expression is tightly controlled by the cyclic AMP receptor protein, CRP. This represents a new link between porin repression and nutrient availability that is likely to be widely conserved among enterobacteria.
    Keywords: Genetic Research -- Genetic Aspects ; Genetic Research -- Physiological Aspects ; Bacterial Genetics -- Genetic Aspects ; Bacterial Genetics -- Physiological Aspects ; Cyclic Adenosine Monophosphate -- Genetic Aspects ; Cyclic Adenosine Monophosphate -- Physiological Aspects ; Salmonella -- Genetic Aspects ; Salmonella -- Physiological Aspects ; Rna -- Genetic Aspects ; Rna -- Physiological Aspects ; Porins -- Genetic Aspects ; Porins -- Physiological Aspects;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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  • 10
    In: Molecular Microbiology, March 2009, Vol.71(5), pp.1228-1238
    Description: The alternative sigma factor σ is activated by unfolded outer membrane proteins (OMPs) and plays an essential role in pathogenesis. The canonical pathway of σ activation in response to envelope stress involves sequential proteolysis of the anti‐sigma factor RseA by the PDZ proteases DegS and RseP. Here we show that σ in sv. Typhimurium can also be activated by acid stress. A σ‐deficient mutant exhibits increased susceptibility to acid pH and reduced survival in an acidified phagosomal vacuole. Acid activation of σ‐dependent gene expression is independent of the unfolded OMP signal or the DegS protease but requires processing of RseA by RseP. The RseP PDZ domain is indispensable for acid induction, suggesting that acid stress may disrupt an inhibitory interaction between RseA and the RseP PDZ domain to allow RseA proteolysis in the absence of antecedent action of DegS. These observations demonstrate a novel environmental stimulus and activation pathway for the σ regulon that appear to be critically important during –host cell interactions.
    Keywords: Membrane Proteins ; Proteolysis ; Salmonella ; Iron Compounds ; Gene Expression ; Proteases;
    ISSN: 0950-382X
    E-ISSN: 1365-2958
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