Carbon Storage Regulator A (csrA) Gene Regulates Motility and Growth of Bacillus licheniformis in the Presence of Hydrocarbons

ABSTRACTCarbon storage regulator A ofBorrelia burgdorferi(CsrABb) contributes to vertebrate host-specific adaptation by modulating activation of the Rrp2-RpoN-RpoS pathway and is critical for infectivity. We hypothesized that the functions of CsrABbare dependent on environmental signals and on select residues. We analyzed the phenotype ofcsrABbdeletion and site-specific mutants to determine the conserved and pathogen-specific attributes of CsrABb. Levels of phosphate acetyltransferase (Pta) involved in conversion of acetyl phosphate to acetyl-coenzyme A (acetyl-CoA) and posttranscriptionally regulated by CsrABbin thecsrABbmutant were reduced from or similar to those in the control strains under unfed- or fed-tick conditions, respectively. Increased levels of supplemental acetate restored vertebrate host-responsive determinants in thecsrABbmutant to parental levels, indicating that both the levels of CsrABband the acetyl phosphate and acetyl-CoA balance contribute to the activation of the Rrp2-RpoN-RpoS pathway. Site-specific replacement of 8 key residues of CsrABb(8S) with alanines resulted in increased levels of CsrABband reduced levels of Pta and acetyl-CoA, while levels of RpoS, BosR, and other members ofrpoSregulon were elevated. Truncation of 7 amino acids at the C terminus of CsrABb(7D) resulted in reducedcsrABbtranscripts and posttranscriptionally reduced levels of FliW located upstream of CsrABb. Electrophoretic mobility shift assays revealed increased binding of 8S mutant protein to the CsrA binding box upstream ofptacompared to the parental and 7D truncated protein. Two CsrABbbinding sites were also identified upstream offliWwithin theflgKcoding sequence. These observations reveal conserved and unique functions of CsrABbthat regulate adaptive gene expression inB. burgdorferi.

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Solution Structure of the Carbon Storage Regulator Protein CsrA from Escherichia coli

Journal of Bacteriology ◽

10.1128/jb.187.10.3496-3501.2005 ◽

2005 ◽

Vol 187 (10) ◽

pp. 3496-3501 ◽

Cited By ~ 62

Author(s):

Pablo Gutiérrez ◽

Yan Li ◽

Michael J. Osborne ◽

Ekaterina Pomerantseva ◽

Qian Liu ◽

...

Keyword(s):

Carbon Storage ◽

Solution Structure ◽

Rna Binding ◽

Binding Motif ◽

Binding Properties ◽

Nmr Titration ◽

C Terminus ◽

Regulator Protein ◽

Carbon Storage Regulator ◽

Α Helix

ABSTRACT The carbon storage regulator A (CsrA) is a protein responsible for the repression of a variety of stationary-phase genes in bacteria. In this work, we describe the nuclear magnetic resonance (NMR)-based structure of the CsrA dimer and its RNA-binding properties. CsrA is a dimer of two identical subunits, each composed of five strands, a small α-helix and a flexible C terminus. NMR titration experiments suggest that the β1-β2 and β3-β4 loops and the C-terminal helix are important elements in RNA binding. Even though the β3-β4 loop contains a highly conserved RNA-binding motif, GxxG, typical of KH domains, our structure excludes CsrA from being a member of this protein family, as previously suggested. A mechanism for the recognition of mRNAs downregulated by CsrA is proposed.

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Carbon storage regulator CsrA plays important roles in multiple virulence-associated processes of Clostridium difficile

Microbial Pathogenesis ◽

10.1016/j.micpath.2018.05.052 ◽

2018 ◽

Vol 121 ◽

pp. 303-309 ◽

Cited By ~ 2

Author(s):

Huawei Gu ◽

Haonan Qi ◽

Shuyi Chen ◽

Kan Shi ◽

Haiying Wang ◽

...

Keyword(s):

Clostridium Difficile ◽

Carbon Storage ◽

Carbon Storage Regulator

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Carbon storage regulator A (CsrABb) is a repressor of Borrelia burgdorferi flagellin protein FlaB

Molecular Microbiology ◽

10.1111/j.1365-2958.2011.07853.x ◽

2011 ◽

Vol 82 (4) ◽

pp. 851-864 ◽

Cited By ~ 41

Author(s):

Ching Wooen Sze ◽

Dustin R. Morado ◽

Jun Liu ◽

Nyles W. Charon ◽

Hongbin Xu ◽

...

Keyword(s):

Borrelia Burgdorferi ◽

Carbon Storage ◽

Carbon Storage Regulator

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Carbon Storage Regulator A Contributes to the Virulence of Haemophilus ducreyi in Humans by Multiple Mechanisms

Infection and Immunity ◽

10.1128/iai.01239-12 ◽

2012 ◽

Vol 81 (2) ◽

pp. 608-617 ◽

Cited By ~ 13

Author(s):

Dharanesh Gangaiah ◽

Wei Li ◽

Kate R. Fortney ◽

Diane M. Janowicz ◽

Sheila Ellinger ◽

...

Keyword(s):

Oxidative Stress ◽

Stationary Phase ◽

Carbon Storage ◽

Stress Responses ◽

Haemophilus Ducreyi ◽

Content Type ◽

Enhanced Sensitivity ◽

Significant Survival ◽

Carbon Storage Regulator ◽

Mutant Phenotypes

ABSTRACTThe carbon storage regulator A (CsrA) controls a wide variety of bacterial processes, including metabolism, adherence, stress responses, and virulence.Haemophilus ducreyi, the causative agent of chancroid, harbors a homolog ofcsrA. Here, we generated an unmarked, in-frame deletion mutant ofcsrAto assess its contribution toH. ducreyipathogenesis. In human inoculation experiments, thecsrAmutant was partially attenuated for pustule formation compared to its parent. Deletion ofcsrAresulted in decreased adherence ofH. ducreyito human foreskin fibroblasts (HFF); Flp1 and Flp2, the determinants ofH. ducreyiadherence to HFF cells, were downregulated in thecsrAmutant. Compared to its parent, thecsrAmutant had a significantly reduced ability to tolerate oxidative stress and heat shock. The enhanced sensitivity of the mutant to oxidative stress was more pronounced in bacteria grown to stationary phase compared to that in bacteria grown to mid-log phase. ThecsrAmutant also had a significant survival defect within human macrophages when the bacteria were grown to stationary phase but not to mid-log phase. Complementation intranspartially or fully restored the mutant phenotypes. These data suggest that CsrA contributes to virulence by multiple mechanisms and that these contributions may be more profound in bacterial cell populations that are not rapidly dividing in the human host.

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The Carbon Storage Regulator (Csr) System Exerts a Nutrient-Specific Control over Central Metabolism in Escherichia coli Strain Nissle 1917

PLoS ONE ◽

10.1371/journal.pone.0066386 ◽

2013 ◽

Vol 8 (6) ◽

pp. e66386 ◽

Cited By ~ 38

Author(s):

Olga Revelles ◽

Pierre Millard ◽

Jean-Philippe Nougayrède ◽

Ulrich Dobrindt ◽

Eric Oswald ◽

...

Keyword(s):

Escherichia Coli ◽

Carbon Storage ◽

Coli Strain ◽

Central Metabolism ◽

Carbon Storage Regulator ◽

Specific Control

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CRP-Mediated Carbon Catabolite Regulation of Yersinia pestis Biofilm Formation Is Enhanced by the Carbon Storage Regulator Protein, CsrA

PLoS ONE ◽

10.1371/journal.pone.0135481 ◽

2015 ◽

Vol 10 (8) ◽

pp. e0135481 ◽

Cited By ~ 23

Author(s):

Stephan P. Willias ◽

Sadhana Chauhan ◽

Chien-Chi Lo ◽

Patrick S. G. Chain ◽

Vladimir L. Motin

Keyword(s):

Yersinia Pestis ◽

Carbon Storage ◽

Biofilm Formation ◽

Regulator Protein ◽

Carbon Storage Regulator ◽

Carbon Catabolite Regulation ◽

Catabolite Regulation

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Inactivation ofbb0184, Which Encodes Carbon Storage Regulator A, Represses the Infectivity ofBorrelia burgdorferi

Infection and Immunity ◽

10.1128/iai.00871-10 ◽

2010 ◽

Vol 79 (3) ◽

pp. 1270-1279 ◽

Cited By ~ 42

Author(s):

Ching Wooen Sze ◽

Chunhao Li

Keyword(s):

Carbon Storage ◽

Associated Factors ◽

Animal Studies ◽

Acetate Metabolism ◽

Acetyl Phosphate ◽

Wild Type ◽

Acetyl Coa ◽

Acetyl Coenzyme A ◽

Carbon Storage Regulator ◽

Subsequent Activation

ABSTRACTThe genome ofBorrelia burgdorferi, the Lyme disease spirochete, encodes a homolog (thebb0184gene product) of the carbon storage regulator A protein (CsrABb); recent studies reported that CsrABbis involved in the regulation of several infectivity factors ofB. burgdorferi. However, the mechanism involved remains unknown. In this report, acsrABbmutant was constructed and complemented in an infectious B31A3 strain. Subsequent animal studies showed that the mutant failed to establish an infection in mice, highlighting that CsrABbis required for the infectivity ofB. burgdorferi. Western blot analyses revealed that the virulence-associated factors OspC, DbpB, and DbpA were attenuated in thecsrABbmutant. The Rrp2-RpoN-RpoS pathway (σ54-σSsigma factor cascade) is a central regulon that governs the expression ofospC,dbpB, anddbpA. Further analyses found that the level of RpoS was significantly decreased in the mutant, while the level of Rrp2 remained unchanged. A recent study reported that the overexpression of BB0589, a phosphate acetyl-transferase (Pta) that converts acetyl-phosphate to acetyl-coenzyme A (CoA), led to the inhibition of RpoS and OspC expression, suggesting that acetyl-phosphate is an activator of Rrp2. Along with this report, we found that CsrABbbinds to the leader sequence of thebb0589transcript and that the intracellular level of acetyl-CoA in thecsrABbmutant was significantly increased compared to that of the wild type, suggesting that more acetyl-phosphate was being converted to acetyl-CoA in the mutant. Collectively, these results suggest that CsrABbmay influence the infectivity ofB. burgdorferivia regulation of acetate metabolism and subsequent activation of the Rrp2-RpoN-RpoS pathway.

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Glucose-Specific Enzyme IIA Has Unique Binding Partners in The Vibrio cholerae Biofilm

mBio ◽

10.1128/mbio.00228-12 ◽

2012 ◽

Vol 3 (6) ◽

Cited By ~ 22

Author(s):

Bradley S. Pickering ◽

Daniel R. Smith ◽

Paula I. Watnick

Keyword(s):

Adenylate Cyclase ◽

Vibrio Cholerae ◽

Carbon Storage ◽

Biofilm Formation ◽

Specific Enzyme ◽

Microbial Physiology ◽

Content Type ◽

Multiprotein Complexes ◽

Binding Partners ◽

Carbon Storage Regulator

ABSTRACTGlucose-specific enzyme IIA (EIIAGlc) is a central regulator of bacterial metabolism and an intermediate in the phosphoenolpyruvate phosphotransferase system (PTS), a conserved phosphotransfer cascade that controls carbohydrate transport. We previously reported that EIIAGlcactivates transcription of the genes required forVibrio choleraebiofilm formation. While EIIAGlcmodulates the function of many proteins through a direct interaction, none of the known regulatory binding partners of EIIAGlcactivates biofilm formation. Therefore, we used tandem affinity purification (TAP) to compare binding partners of EIIAGlcin both planktonic and biofilm cells. A surprising number of novel EIIAGlcbinding partners were identified predominantly under one condition or the other. Studies of planktonic cells revealed established partners of EIIAGlc, such as adenylate cyclase and glycerol kinase. In biofilms, MshH, a homolog ofEscherichia coliCsrD, was found to be a dominant binding partner of EIIAGlc. Further studies revealed that MshH inhibits biofilm formation. This function was independent of the Carbon storage regulator (Csr) pathway and dependent on EIIAGlc. To explore the existence of multiprotein complexes centered on EIIAGlc, we also affinity purified the binding partners of adenylate cyclase from biofilm cells. In addition to EIIAGlc, this analysis yielded many of the same proteins that copurified with EIIAGlc. We hypothesize that EIIAGlcserves as a hub for multiprotein complexes and furthermore that these complexes may provide a mechanism for competitive and cooperative interactions between binding partners.IMPORTANCEEIIAGlcis a global regulator of microbial physiology that acts through direct interactions with other proteins. This work represents the first demonstration that the protein partners of EIIAGlcare distinct in the microbial biofilm. Furthermore, it provides the first evidence that EIIAGlcmay exist in multiprotein complexes with its partners, setting the stage for an investigation of how the multiple partners of EIIAGlcinfluence one another. Last, it provides a connection between the phosphoenolpyruvate phosphotransferase (PTS) and Csr (Carbon storage regulator) regulatory systems. This work increases our understanding of the complexity of regulation by EIIAGlcand provides a link between the PTS and Csr networks, two global regulatory cascades that influence microbial physiology.

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