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  • 1
    Online Resource
    Online Resource
    Cold Spring Harbor Laboratory ; 2011
    In:  RNA Vol. 17, No. 6 ( 2011-06), p. 1172-1189
    In: RNA, Cold Spring Harbor Laboratory, Vol. 17, No. 6 ( 2011-06), p. 1172-1189
    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.
    Type of Medium: Online Resource
    ISSN: 1355-8382 , 1469-9001
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2011
    detail.hit.zdb_id: 1475737-0
    SSG: 12
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  • 2
    Online Resource
    Online Resource
    Cold Spring Harbor Laboratory ; 2015
    In:  RNA Vol. 21, No. 4 ( 2015-04), p. 511-512
    In: RNA, Cold Spring Harbor Laboratory, Vol. 21, No. 4 ( 2015-04), p. 511-512
    Type of Medium: Online Resource
    ISSN: 1355-8382 , 1469-9001
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2015
    detail.hit.zdb_id: 1475737-0
    SSG: 12
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  • 3
    Online Resource
    Online Resource
    Cold Spring Harbor Laboratory ; 2023
    In:  Genes & Development Vol. 37, No. 1-2 ( 2023-01-01), p. 27-29
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 37, No. 1-2 ( 2023-01-01), p. 27-29
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    RVK:
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2023
    detail.hit.zdb_id: 1467414-2
    SSG: 12
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  • 4
    Online Resource
    Online Resource
    Cold Spring Harbor Laboratory ; 2001
    In:  Genes & Development Vol. 15, No. 5 ( 2001-03-01), p. 627-637
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 15, No. 5 ( 2001-03-01), p. 627-637
    Abstract: The ς S subunit of Escherichia coli RNA polymerase regulates the expression of stationary phase and stress response genes. Control over ς S activity is exercised in part by regulated degradation of ς S . In vivo, degradation requires the ClpXP protease together with RssB, a protein homologous to response regulator proteins. Using purified components, we reconstructed the degradation of ς S in vitro and demonstrate a direct role for RssB in delivering ς S to ClpXP. RssB greatly stimulates ς S degradation by ClpXP. Acetyl phosphate, which phosphorylates RssB, is required. RssB participates in multiple rounds of ς S degradation, demonstrating its catalytic role. RssB promotes ς S degradation specifically; it does not affect degradation of other ClpXP substrates or other proteins not normally degraded by ClpXP. ς S and RssB form a stable complex in the presence of acetyl phosphate, and together they form a ternary complex with ClpX that is stabilized by ATP[γ-S]. Alone, neither ς S nor RssB binds ClpX with high affinity. When ClpP is present, a larger ς S –RssB–ClpXP complex forms. The complex degrades ς S and releases RssB from ClpXP in an ATP-dependent reaction. Our results illuminate an important mechanism for regulated protein turnover in which a unique targeting protein, whose own activity is regulated through specific signaling pathways, catalyzes the delivery of a specific substrate to a specific protease.
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    RVK:
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2001
    detail.hit.zdb_id: 1467414-2
    SSG: 12
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  • 5
    Online Resource
    Online Resource
    Cold Spring Harbor Laboratory ; 2009
    In:  Genes & Development Vol. 23, No. 17 ( 2009-09-01), p. 1981-1985
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 23, No. 17 ( 2009-09-01), p. 1981-1985
    Abstract: Many bacterial small regulatory RNAs (sRNAs) pair with mRNA targets, stimulating or inhibiting mRNA stability and/or translation. Regulation of these sRNAs is usually due to tight transcriptional regulation of synthesis.In this issue of Genes & Development and a related paper in Molecular Microbiology , Figueroa-Bossi and colleagues (pp. 2004–2015) and Overgaard and colleagues report a novel regulatory mechanism in which induction of a competing mRNA acts to titrate away the sRNA, allowing expression of an otherwise strongly inhibited target gene.
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    RVK:
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2009
    detail.hit.zdb_id: 1467414-2
    SSG: 12
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  • 6
    Online Resource
    Online Resource
    Cold Spring Harbor Laboratory ; 2006
    In:  Genes & Development Vol. 20, No. 17 ( 2006-09-01), p. 2338-2348
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 20, No. 17 ( 2006-09-01), p. 2338-2348
    Abstract: MicF, one of the first chromosomally encoded regulatory small RNAs (sRNAs) to be discovered, was found to modulate the expression of OmpF, an abundant outer membrane protein. Several recent papers have now shown that this is not an isolated case. At least five other sRNAs also regulate the synthesis of outer membrane porins, and additional sRNAs modulate the expression of other outer membrane proteins. Here we review what is known about these sRNAs and discuss the implications of this regulation.
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    RVK:
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2006
    detail.hit.zdb_id: 1467414-2
    SSG: 12
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  • 7
    Online Resource
    Online Resource
    Cold Spring Harbor Laboratory ; 2003
    In:  Genes & Development Vol. 17, No. 19 ( 2003-10-01), p. 2374-2383
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 17, No. 19 ( 2003-10-01), p. 2374-2383
    Abstract: RyhB is a small antisense regulatory RNA that is repressed by the Fur repressor and negatively regulates at least six mRNAs encoding Fe-binding or Fe-storage proteins in Escherichia coli . When Fe is limiting, RyhB levels rise, and target mRNAs are rapidly degraded. RyhB is very stable when measured after treatment of cells with the transcription inhibitor rifampicin, but is unstable when overall mRNA transcription continues. We propose that RyhB turnover is coupled to and dependent on pairing with the target mRNAs. Degradation of both mRNA targets and RyhB is dependent on RNase E and is slowed in degradosome mutants. RyhB requires the RNA chaperone Hfq. In the absence of Hfq, RyhB is unstable, even when general transcription is inhibited; degradation is dependent upon RNase E. Hfq and RNase E bind similar sites on the RNA; pairing may allow loss of Hfq and access by RNase E. Two other Hfq-dependent small RNAs, DsrA and OxyS, are also stable when overall transcription is off, and unstable when it is not, suggesting that they, too, are degraded when their target mRNAs are available for pairing. Thus, this large class of regulatory RNAs share an unexpected intrinsic mechanism for shutting off their action.
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    RVK:
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2003
    detail.hit.zdb_id: 1467414-2
    SSG: 12
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  • 8
    Online Resource
    Online Resource
    Cold Spring Harbor Laboratory ; 2006
    In:  Genes & Development Vol. 20, No. 7 ( 2006-04-01), p. 884-897
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 20, No. 7 ( 2006-04-01), p. 884-897
    Abstract: The σ S subunit of Escherichia coli RNA polymerase regulates the expression of stationary phase and stress response genes. σ S is highly unstable in exponentially growing cells, whereas its stability increases dramatically upon starvation or under certain stress conditions. The degradation of σ S is controlled by the phosphorylatable adaptor protein RssB and the ClpXP protease. RssB specifically directs σ S to ClpXP. An unanswered question is how RssB-mediated degradation of σ S is blocked by conditions such as glucose or phosphate starvation. We report here the identification and characterization of a new regulator of σ S stability, IraP (inhibitor of RssB activity during phosphate starvation), that stabilizes σ S both in vivo and in vitro. Deletion of iraP interferes with σ S stabilization during phosphate starvation, but not during carbon starvation, and has a partial effect in stationary phase and nitrogen starvation. IraP interferes with RssB-dependent degradation of σ S through a direct protein–protein interaction with RssB. A point mutant of IraP was isolated and found to be defective both for inhibition of σ S degradation and interaction with RssB. Our results reveal a novel mechanism of regulation of σ S stability through the regulation of RssB activity and identify IraP as a member of a new class of regulators, the anti-adaptor proteins.
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    RVK:
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2006
    detail.hit.zdb_id: 1467414-2
    SSG: 12
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  • 9
    Online Resource
    Online Resource
    Cold Spring Harbor Laboratory ; 2017
    In:  Genes & Development Vol. 31, No. 13 ( 2017-07-01), p. 1382-1395
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 31, No. 13 ( 2017-07-01), p. 1382-1395
    Abstract: Mismatch repair (MMR) is a conserved mechanism exploited by cells to correct DNA replication errors both in growing cells and under nongrowing conditions. Hfq (host factor for RNA bacteriophage Qβ replication), a bacterial Lsm family RNA-binding protein, chaperones RNA–RNA interactions between regulatory small RNAs (sRNAs) and target messenger RNAs (mRNAs), leading to alterations of mRNA translation and/or stability. Hfq has been reported to post-transcriptionally repress the DNA MMR gene mutS in stationary phase, possibly limiting MMR to allow increased mutagenesis. Here we report that Hfq deploys dual mechanisms to control mutS expression. First, Hfq binds directly to an (AAN) 3 motif within the mutS 5′ untranslated region (UTR), repressing translation in the absence of sRNA partners both in vivo and in vitro. Second, Hfq acts in a canonical pathway, promoting base-pairing of ArcZ sRNA with the mutS leader to inhibit translation. Most importantly, using pathway-specific mutS chromosomal alleles that specifically abrogate either regulatory pathway or both, we demonstrate that tight control of MutS levels in stationary phase contributes to stress-induced mutagenesis. By interacting with the mutS leader, Hfq serves as a critical switch that modulates bacteria from high-fidelity DNA replication to stress-induced mutagenesis.
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    RVK:
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2017
    detail.hit.zdb_id: 1467414-2
    SSG: 12
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  • 10
    Online Resource
    Online Resource
    Cold Spring Harbor Laboratory ; 2002
    In:  Genes & Development Vol. 16, No. 22 ( 2002-11-15), p. 2829-2842
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 16, No. 22 ( 2002-11-15), p. 2829-2842
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    RVK:
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2002
    detail.hit.zdb_id: 1467414-2
    SSG: 12
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