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      A search for small noncoding RNAs in Staphylococcus aureus reveals a conserved sequence motif for regulation

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          Abstract

          Bioinformatic analysis of the intergenic regions of Staphylococcus aureus predicted multiple regulatory regions. From this analysis, we characterized 11 novel noncoding RNAs (RsaA‐K) that are expressed in several S. aureus strains under different experimental conditions. Many of them accumulate in the late-exponential phase of growth. All ncRNAs are stable and their expression is Hfq-independent. The transcription of several of them is regulated by the alternative sigma B factor (RsaA, D and F) while the expression of RsaE is agrA-dependent. Six of these ncRNAs are specific to S. aureus, four are conserved in other Staphylococci, and RsaE is also present in Bacillaceae. Transcriptomic and proteomic analysis indicated that RsaE regulates the synthesis of proteins involved in various metabolic pathways. Phylogenetic analysis combined with RNA structure probing, searches for RsaE‐mRNA base pairing, and toeprinting assays indicate that a conserved and unpaired UCCC sequence motif of RsaE binds to target mRNAs and prevents the formation of the ribosomal initiation complex. This study unexpectedly shows that most of the novel ncRNAs carry the conserved C−rich motif, suggesting that they are members of a class of ncRNAs that target mRNAs by a shared mechanism.

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          Most cited references94

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          Vienna RNA secondary structure server.

          The Vienna RNA secondary structure server provides a web interface to the most frequently used functions of the Vienna RNA software package for the analysis of RNA secondary structures. It currently offers prediction of secondary structure from a single sequence, prediction of the consensus secondary structure for a set of aligned sequences and the design of sequences that will fold into a predefined structure. All three services can be accessed via the Vienna RNA web server at http://rna.tbi.univie.ac.at/.
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            Regulatory RNAs in bacteria.

            Bacteria possess numerous and diverse means of gene regulation using RNA molecules, including mRNA leaders that affect expression in cis, small RNAs that bind to proteins or base pair with target RNAs, and CRISPR RNAs that inhibit the uptake of foreign DNA. Although examples of RNA regulators have been known for decades in bacteria, we are only now coming to a full appreciation of their importance and prevalence. Here, we review the known mechanisms and roles of regulatory RNAs, highlight emerging themes, and discuss remaining questions.
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              Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.

              A discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) system for the separation of proteins in the range from 1 to 100 kDa is described. Tricine, used as the trailing ion, allows a resolution of small proteins at lower acrylamide concentrations than in glycine-SDS-PAGE systems. A superior resolution of proteins, especially in the range between 5 and 20 kDa, is achieved without the necessity to use urea. Proteins above 30 kDa are already destacked within the sample gel. Thus a smooth passage of these proteins from sample to separating gel is warranted and overloading effects are reduced. This is of special importance when large amounts of protein are to be loaded onto preparative gels. The omission of glycine and urea prevents disturbances which might occur in the course of subsequent amino acid sequencing.
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                Author and article information

                Journal
                Nucleic Acids Res
                Nucleic Acids Res
                nar
                nar
                Nucleic Acids Research
                Oxford University Press
                0305-1048
                1362-4962
                November 2009
                November 2009
                28 September 2009
                28 September 2009
                : 37
                : 21
                : 7239-7257
                Affiliations
                1Architecture et Réactivité de l’ARN, Université de Strasbourg, CNRS, IBMC, 15 rue René Descartes, F-67084 Strasbourg, 2INRA, UR 875, F-31320 Castanet-Tolosan, 3INSERM U851, Centre National de Référence des Staphylocoques, Université de Lyon I, F-69008 Lyon, France and 4Genomic Research Laboratory, Service of Infectious Diseases, University of Geneva Hospitals, Rue Micheli-du-Crest, 24, CH-1211 Geneva 14, Switzerland
                Author notes
                *To whom correspondence should be addressed. Tel: 33 3 88417051; Fax: 33 3 88602218; Email: p.romby@ 123456ibmc.u-strasbg.fr
                Correspondence may also be addressed to Christine Gaspin. Email: gaspin@ 123456toulouse.inra.fr
                Correspondence may also be addressed to François Vandenesch. Email: denesch@ 123456univ-lyon1.fr

                Present address: Thomas Geissmann, INSERM U851, Centre National de Référence des Staphylocoques, Université de Lyon I, F-69008 Lyon, France.

                Article
                gkp668
                10.1093/nar/gkp668
                2790875
                19786493
                8cb3860d-aa7f-4b89-a0e0-81da57148589
                © The Author(s) 2009. Published by Oxford University Press.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 22 May 2009
                : 24 July 2009
                : 29 July 2009
                Categories
                RNA

                Genetics
                Genetics

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