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  • RNA, Small Untranslated  (30)
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  • 1
    Language: English
    In: Proceedings of the National Academy of Sciences of the United States of America, 02 August 2011, Vol.108(31), pp.12875-80
    Description: The Escherichia coli σ(E) envelope stress response monitors and repairs the outer membrane, a function central to the life of Gram-negative bacteria. The σ(E) stress response was characterized as a single-tier activation network comprised of ~100 genes, including the MicA and RybB noncoding sRNAs. These highly expressed sRNAs were thought to carry out the specialized function of halting de novo synthesis of several abundant porins when envelope homeostasis was perturbed. Using a systematic target profiling and validation approach we discovered that MicA and RybB are each global mRNA repressors of both distinct and shared targets, and that the two sRNAs constitute a posttranscriptional repression arm whose regulatory scope rivals that of the protein-based σ(E) activation arm. Intriguingly, porin mRNAs constitute only ~1/3 of all targets and new nonporin targets predict roles for MicA and RybB in crosstalk with other regulatory responses. This work also provides an example of evolutionarily unrelated sRNAs that are coinduced and bind the same targets, but at different sites. Our finding that expression of either MicA or RybB sRNA protects the cell from the loss of viability experienced when σ(E) activity is inadequate illustrates the importance of the posttranscriptional repression arm of the response. σ(E) is a paradigm of a single-tier stress response with a clear division of labor in which highly expressed noncoding RNAs (MicA, RybB) endow a transcriptional factor intrinsically restricted to gene activation (σ(E)) with the opposite repressor function.
    Keywords: Escherichia Coli Proteins -- Genetics ; RNA, Small Untranslated -- Genetics ; Regulon -- Genetics ; Sigma Factor -- Genetics
    ISSN: 00278424
    E-ISSN: 1091-6490
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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
    Language: English
    In: Philosophical transactions of the Royal Society of London. Series B, Biological sciences, 05 November 2016, Vol.371(1707)
    Description: Infection is a complicated balance, with both pathogen and host struggling to tilt the result in their favour. Bacterial infection biology has relied on forward genetics for many of its advances, defining phenotype in terms of replication in model systems. However, many known virulence factors fail to produce robust phenotypes, particularly in the systems most amenable to genetic manipulation, such as cell-culture models. This has particularly been limiting for the study of the bacterial regulatory small RNAs (sRNAs) in infection. We argue that new sequencing-based technologies can work around this problem by providing a 'molecular phenotype', defined in terms of the specific transcriptional dysregulation in the infection system induced by gene deletion. We illustrate this using the example of our recent study of the PinT sRNA using dual RNA-seq, that is, simultaneous RNA sequencing of host and pathogen during infection. We additionally discuss how other high-throughput technologies, in particular genetic interaction mapping using transposon insertion sequencing, may be used to further dissect molecular phenotypes. We propose a strategy for how high-throughput technologies can be integrated in the study of non-coding regulators as well as bacterial virulence factors, enhancing our ability to rapidly generate hypotheses with regards to their function.This article is part of the themed issue 'The new bacteriology'.
    Keywords: Pint ; Tn-Seq ; Dual RNA-Seq ; Host–Pathogen Interaction ; Infection ; Small Non-Coding RNA ; Chromosome Mapping -- Methods ; High-Throughput Nucleotide Sequencing -- Methods ; RNA, Bacterial -- Genetics ; RNA, Small Untranslated -- Genetics ; Sequence Analysis, RNA -- Methods
    ISSN: 09628436
    E-ISSN: 1471-2970
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  • 4
    In: EMBO Journal, 03 June 2015, Vol.34(11), pp.1478-1492
    Description: There is an expanding list of examples by which one can posttranscriptionally influence the expression of others. This can involve sponges that sequester regulatory s of s in the same regulon, but the underlying molecular mechanism of such cross talk remains little understood. Here, we report sponge‐mediated cross talk in the posttranscriptional network of GcvB, a conserved Hfq‐dependent small with one of the largest regulons known in bacteria. We show that decay from the locus encoding an amino acid transporter generates a stable fragment (SroC) that base‐pairs with GcvB. This interaction triggers the degradation of GcvB by ase E, alleviating the GcvB‐mediated repression of other amino acid‐related transport and metabolic genes. Intriguingly, since the itself is a target of GcvB, the SroC sponge seems to enable both an internal feed‐forward loop to activate its parental in and activation of many ‐encoded s in the same pathway. Disabling this cross talk affects bacterial growth when peptides are the sole carbon and nitrogen sources. Decay of the bacterial GcvB , which keeps it from regulating its targets, is triggered by a 3′‐‐derived fragment from a target . This ability of s to compete for regulatory interaction presents a new mode of cross talk in bacteria. . Decay of the bacterial GcvB s, which keeps it from regulating its m targets, is triggered by a 3′‐‐derived fragment from a target m. This ability of ms to compete for regulatory interaction presents a new mode of cross talk in bacteria.
    Keywords: G Cv B ; H Fq ; Noncoding Rna ; Rn Ase E ; S Ro C
    ISSN: 0261-4189
    E-ISSN: 1460-2075
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  • 5
    Language: English
    In: Cell, 11 April 2013, Vol.153(2), pp.426-437
    Description: Glucose homeostasis is strictly controlled in all domains of life. Bacteria that are unable to balance intracellular sugar levels and deal with potentially toxic phosphosugars cease growth and risk being outcompeted. Here, we identify the conserved haloacid dehalogenase (HAD)-like enzyme YigL as the previously hypothesized phosphatase for detoxification of phosphosugars and reveal that its synthesis is activated by an Hfq-dependent small RNA in . We show that the glucose-6-P-responsive small RNA SgrS activates YigL synthesis in a translation-independent fashion by the selective stabilization of a decay intermediate of the dicistronic messenger RNA (mRNA). Intriguingly, the major endoribonuclease RNase E, previously known to function together with small RNAs to degrade mRNA targets, is also essential for this process of mRNA activation. The exploitation of and targeted interference with regular RNA turnover described here may constitute a general route for small RNAs to rapidly activate both coding and noncoding genes. ► The bacterial small RNA SgrS posttranscriptionally activates the synthesis of YigL ► YigL is the previously hypothesized phosphatase that prevents phosphosugar toxicity ► SgrS activates yigL by a translation-independent mRNA-stabilization mechanism ► SgrS stabilizes an intermediate in the yigL mRNA decay pathway YigL, a long-sought bacterial phosphatase, regulates glucose-6-phosphate levels. A small regulatory RNA upregulates YigL synthesis by base pairing with the coding sequence of the preceding gene to interfere with endonucleolytic yigL mRNA decay.
    Keywords: Biology
    ISSN: 0092-8674
    E-ISSN: 1097-4172
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  • 6
    In: EMBO Journal, 17 October 2012, Vol.31(20), pp.4005-4019
    Description: The small RNAs associated with the protein Hfq constitute one of the largest classes of post‐transcriptional regulators known to date. Most previously investigated members of this class are encoded by conserved free‐standing genes. Here, deep sequencing of Hfq‐bound transcripts from multiple stages of growth of revealed a plethora of new small RNA species from within mRNA loci, including DapZ, which overlaps with the 3′ region of the biosynthetic gene, . Synthesis of the DapZ small RNA is independent of DapB protein synthesis, and is controlled by HilD, the master regulator of invasion genes. DapZ carries a short G/U‐rich domain similar to that of the globally acting GcvB small RNA, and uses GcvB‐like seed pairing to repress translation of the major ABC transporters, DppA and OppA. This exemplifies double functional output from an mRNA locus by the production of both a protein and an Hfq‐dependent ‐acting RNA. Our atlas of Hfq targets suggests that the 3′ regions of mRNA genes constitute a rich reservoir that provides the Hfq network with new regulatory small RNAs. Deep sequencing of Hfq‐binding RNAs isolated from at different growth stages reveals that the 3′ UTR of bacterial mRNAs are a rich source of regulatory small RNAs which modulate gene expression in trans.
    Keywords: Abc Transporter ; Dapz ; Gcvb ; Hfq ; 3′ Utr
    ISSN: 0261-4189
    E-ISSN: 1460-2075
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  • 7
    In: EMBO Journal, 13 April 2017, Vol.36(8), pp.1029-1045
    Description: Research into post‐transcriptional control of mRNAs by small noncoding RNAs (sRNAs) in the model bacteria and has mainly focused on sRNAs that associate with the RNA chaperone Hfq. However, the recent discovery of the protein ProQ as a common binding partner that stabilizes a distinct large class of structured sRNAs suggests that additional RNA regulons exist in these organisms. The cellular functions and molecular mechanisms of these new ProQ‐dependent sRNAs are largely unknown. Here, we report in Typhimurium the mode‐of‐action of RaiZ, a ProQ‐dependent sRNA that is made from the 3′ end of the mRNA encoding ribosome‐inactivating protein RaiA. We show that RaiZ is a base‐pairing sRNA that represses in the mRNA of histone‐like protein HU‐α. RaiZ forms an RNA duplex with the ribosome‐binding site of mRNA, facilitated by ProQ, to prevent 30S ribosome loading and protein synthesis of HU‐α. Similarities and differences between ProQ‐ and Hfq‐mediated regulation will be discussed. The enterobacterial sRNA RaiZ functions independent of the Hfq RNA chaperone via the recently identified general RNA‐binding protein ProQ. ProQ acts in a dual manner, stabilizing the sRNA and facilitating translational repression of the nucleid protein HU‐α. RaiZ is a small RNA produced by RNase E‐mediated cleavage of the raiA mRNA. RaiZ strongly binds RNA chaperone ProQ, leading to RaiZ stabilization. RaiZ represses translation of the hupA mRNA by base pairing with its ribosome‐binding site. ProQ and RaiZ jointly prevent initiating ribosomes from loading on hupA mRNA. The global RNA‐binding protein ProQ stabilizes bacterial small RNA RaiZ and facilitates translational repression of its target mRNA, thus exemplifying an Hfq‐independent RNA regulon.
    Keywords: Hu‐Α ; Proq ; Raiz ; Small Rna ; Translation Inhibition
    ISSN: 0261-4189
    E-ISSN: 1460-2075
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  • 8
    Language: English
    In: Nucleic acids research, October 2010, Vol.38(19), pp.6637-51
    Description: Post-transcriptional regulatory mechanisms are widespread in bacteria. Interestingly, current published data hint that some of these mechanisms may be non-random with respect to their phylogenetic distribution. Although small, trans-acting regulatory RNAs commonly occur in bacterial genomes, they have been better characterized in Gram-negative bacteria, leaving the impression that they may be less important for Firmicutes. It has been presumed that Gram-positive bacteria, in particular the Firmicutes, are likely to utilize cis-acting regulatory RNAs located within the 5' mRNA leader region more often than trans-acting regulatory RNAs. In this analysis we catalog, by a deep sequencing-based approach, both classes of regulatory RNA candidates for Bacillus subtilis, the model microorganism for Firmicutes. We successfully recover most of the known small RNA regulators while also identifying a greater number of new candidate RNAs. We anticipate these data to be a broadly useful resource for analysis of post-transcriptional regulatory strategies in B. subtilis and other Firmicutes.
    Keywords: Bacillus Subtilis -- Genetics ; RNA, Small Untranslated -- Analysis
    ISSN: 03051048
    E-ISSN: 1362-4962
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  • 9
    Language: English
    In: Proceedings of the National Academy of Sciences of the United States of America, 15 May 2012, Vol.109(20), pp.E1277-86
    Description: More than 50 y of research have provided great insight into the physiology, metabolism, and molecular biology of Salmonella enterica serovar Typhimurium (S. Typhimurium), but important gaps in our knowledge remain. It is clear that a precise choreography of gene expression is required for Salmonella infection, but basic genetic information such as the global locations of transcription start sites (TSSs) has been lacking. We combined three RNA-sequencing techniques and two sequencing platforms to generate a robust picture of transcription in S. Typhimurium. Differential RNA sequencing identified 1,873 TSSs on the chromosome of S. Typhimurium SL1344 and 13% of these TSSs initiated antisense transcripts. Unique findings include the TSSs of the virulence regulators phoP, slyA, and invF. Chromatin immunoprecipitation revealed that RNA polymerase was bound to 70% of the TSSs, and two-thirds of these TSSs were associated with σ(70) (including phoP, slyA, and invF) from which we identified the -10 and -35 motifs of σ(70)-dependent S. Typhimurium gene promoters. Overall, we corrected the location of important genes and discovered 18 times more promoters than identified previously. S. Typhimurium expresses 140 small regulatory RNAs (sRNAs) at early stationary phase, including 60 newly identified sRNAs. Almost half of the experimentally verified sRNAs were found to be unique to the Salmonella genus, and 〈20% were found throughout the Enterobacteriaceae. This description of the transcriptional map of SL1344 advances our understanding of S. Typhimurium, arguably the most important bacterial infection model.
    Keywords: Gene Expression Regulation, Bacterial -- Genetics ; RNA, Small Untranslated -- Genetics ; Regulatory Sequences, Ribonucleic Acid -- Genetics ; Salmonella Typhimurium -- Genetics ; Transcription, Genetic -- Genetics
    ISSN: 00278424
    E-ISSN: 1091-6490
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  • 10
    Language: English
    In: Nucleic acids research, March 2012, Vol.40(5), pp.2020-31
    Description: The Gram-negative plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria (Xcv) is an important model to elucidate the mechanisms involved in the interaction with the host. To gain insight into the transcriptome of the Xcv strain 85-10, we took a differential RNA sequencing (dRNA-seq) approach. Using a novel method to automatically generate comprehensive transcription start site (TSS) maps we report 1421 putative TSSs in the Xcv genome. Genes in Xcv exhibit a poorly conserved -10 promoter element and no consensus Shine-Dalgarno sequence. Moreover, 14% of all mRNAs are leaderless and 13% of them have unusually long 5'-UTRs. Northern blot analyses confirmed 16 intergenic small RNAs and seven cis-encoded antisense RNAs in Xcv. Expression of eight intergenic transcripts was controlled by HrpG and HrpX, key regulators of the Xcv type III secretion system. More detailed characterization identified sX12 as a small RNA that controls virulence of Xcv by affecting the interaction of the pathogen and its host plants. The transcriptional landscape of Xcv is unexpectedly complex, featuring abundant antisense transcripts, alternative TSSs and clade-specific small RNAs.
    Keywords: RNA, Small Untranslated -- Metabolism ; Virulence Factors -- Genetics ; Xanthomonas Campestris -- Genetics
    ISSN: 03051048
    E-ISSN: 1362-4962
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