PMID:21527671: Difference between revisions

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<span style = 'font-size: large'> '''De Lay, N and Gottesman, S'''  (2011) Role of polynucleotide phosphorylase in sRNA function in Escherichia coli.''RNA'' '''17''':1172-89</span>
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|{{TableHeadingColor}} |'''Abstract''' || <protect>In Escherichia coli, many small noncoding regulatory RNAs (sRNAs) post-transcriptionally regulate gene expression by base-pairing to mRNAs in a process that is mediated by the RNA chaperone Hfq. Binding of the sRNA to the mRNA can lead to increased or decreased mRNA stability and/or translation. It is not known if proteins other than Hfq are necessary for this process. In order to identify additional genes required for the post-transcriptional regulation of gene expression by Hfq-dependent sRNAs, we developed a novel combined genetic selection and screen for mutants defective in sRNA regulation. In our combined genetic selection and screen, we isolated hfq mutants and mutants in pnp, encoding polynucleotide phosphorylase (PNPase). We show that loss-of-function mutations in pnp result in a decreased stability of several sRNAs including RyhB, SgrS, and CyaR and also decrease both the negative and positive regulation by sRNAs. The defect in stability of CyaR and in negative and positive regulation are suppressed by deletion mutations in RNase E. Altogether, our results suggest that the lack of sRNA-mediated regulation in the absence of an active form of PNPase is due to the rapid turnover of sRNA resulting from an increase in RNase E activity and/or an increase in access of other ribonucleases to sRNAs.</protect>
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'''No author listed''' (2011) Role of polynucleotide phosphorylase in sRNA function in Escherichia coli. ''RNA'' '''17''':1172-89
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!align=left  |Abstract
*  [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=21527671 PubMed]
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In Escherichia coli, many small noncoding regulatory RNAs (sRNAs) post-transcriptionally regulate gene expression by base-pairing to mRNAs in a process that is mediated by the RNA chaperone Hfq. Binding of the sRNA to the mRNA can lead to increased or decreased mRNA stability and/or translation. It is not known if proteins other than Hfq are necessary for this process. In order to identify additional genes required for the post-transcriptional regulation of gene expression by Hfq-dependent sRNAs, we developed a novel combined genetic selection and screen for mutants defective in sRNA regulation. In our combined genetic selection and screen, we isolated hfq mutants and mutants in pnp, encoding polynucleotide phosphorylase (PNPase). We show that loss-of-function mutations in pnp result in a decreased stability of several sRNAs including RyhB, SgrS, and CyaR and also decrease both the negative and positive regulation by sRNAs. The defect in stability of CyaR and in negative and positive regulation are suppressed by deletion mutations in RNase E. Altogether, our results suggest that the lack of sRNA-mediated regulation in the absence of an active form of PNPase is due to the rapid turnover of sRNA resulting from an increase in RNase E activity and/or an increase in access of other ribonucleases to sRNAs.
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[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=21527671 PubMed] [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3096048 PMC3096048]
Online version:[http://dx.doi.org/10.1261/rna.2531211 10.1261/rna.2531211]
Online version:[http://dx.doi.org/10.1261/rna.2531211 10.1261/rna.2531211]
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!align=left  |Keywords
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Endoribonucleases/genetics; Endoribonucleases/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli/metabolism; Gene Expression Regulation, Bacterial; Host Factor 1 Protein/genetics; Host Factor 1 Protein/metabolism; Polyribonucleotide Nucleotidyltransferase/genetics; Polyribonucleotide Nucleotidyltransferase/metabolism; RNA Stability/genetics; RNA, Bacterial/genetics; RNA, Bacterial/metabolism; RNA, Messenger/metabolism; RNA, Small Untranslated/metabolism
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Revision as of 19:13, 1 September 2013

Citation

No author listed (2011) Role of polynucleotide phosphorylase in sRNA function in Escherichia coli. RNA 17:1172-89

Abstract

In Escherichia coli, many small noncoding regulatory RNAs (sRNAs) post-transcriptionally regulate gene expression by base-pairing to mRNAs in a process that is mediated by the RNA chaperone Hfq. Binding of the sRNA to the mRNA can lead to increased or decreased mRNA stability and/or translation. It is not known if proteins other than Hfq are necessary for this process. In order to identify additional genes required for the post-transcriptional regulation of gene expression by Hfq-dependent sRNAs, we developed a novel combined genetic selection and screen for mutants defective in sRNA regulation. In our combined genetic selection and screen, we isolated hfq mutants and mutants in pnp, encoding polynucleotide phosphorylase (PNPase). We show that loss-of-function mutations in pnp result in a decreased stability of several sRNAs including RyhB, SgrS, and CyaR and also decrease both the negative and positive regulation by sRNAs. The defect in stability of CyaR and in negative and positive regulation are suppressed by deletion mutations in RNase E. Altogether, our results suggest that the lack of sRNA-mediated regulation in the absence of an active form of PNPase is due to the rapid turnover of sRNA resulting from an increase in RNase E activity and/or an increase in access of other ribonucleases to sRNAs.

Links

PubMed PMC3096048 Online version:10.1261/rna.2531211

Keywords

Endoribonucleases/genetics; Endoribonucleases/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli/metabolism; Gene Expression Regulation, Bacterial; Host Factor 1 Protein/genetics; Host Factor 1 Protein/metabolism; Polyribonucleotide Nucleotidyltransferase/genetics; Polyribonucleotide Nucleotidyltransferase/metabolism; RNA Stability/genetics; RNA, Bacterial/genetics; RNA, Bacterial/metabolism; RNA, Messenger/metabolism; RNA, Small Untranslated/metabolism

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