The agr Locus Regulates Virulence and Colonization Genes in Clostridium difficile 027

ABSTRACT The transcriptional regulator AgrA, a member of the LytTR family of proteins, plays a key role in controlling gene expression in some Gram-positive pathogens, including Staphylococcus aureus and Enterococcus faecalis. AgrA is encoded by the agrACDB global regulatory locus, and orthologues are found within the genome of most Clostridium difficile isolates, including the epidemic lineage 027/BI/NAP1. Comparative RNA sequencing of the wild type and otherwise isogenic agrA null mutant derivatives of C. difficile R20291 revealed a network of approximately 75 differentially regulated transcripts at late exponential growth phase, including many genes associated with flagellar assembly and function, such as the major structural subunit, FliC. Other differentially regulated genes include several involved in bis-(3′-5′)-cyclic dimeric GMP (c-di-GMP) synthesis and toxin A expression. C. difficile 027 R20291 agrA mutant derivatives were poorly flagellated and exhibited reduced levels of colonization and relapses in the murine infection model. Thus, the agr locus likely plays a contributory role in the fitness and virulence potential of C. difficile strains in the 027/BI/NAP1 lineage.

[1]  D. Sidote,et al.  Identification of a hydrophobic cleft in the LytTR domain of AgrA as a locus for small molecule interactions that inhibit DNA binding. , 2012, Biochemistry.

[2]  D. Serruto,et al.  Multiple Factors Modulate Biofilm Formation by the Anaerobic Pathogen Clostridium difficile , 2012, Journal of bacteriology.

[3]  M. Pirmohamed,et al.  Emergence and global spread of epidemic healthcare-associated Clostridium difficile , 2012, Nature Genetics.

[4]  Sean R. Eddy,et al.  Rfam 11.0: 10 years of RNA families , 2012, Nucleic Acids Res..

[5]  E. Kuijper,et al.  C. difficile 630Δerm Spo0A Regulates Sporulation, but Does Not Contribute to Toxin Production, by Direct High-Affinity Binding to Target DNA , 2012, PloS one.

[6]  Taane G. Clark,et al.  Targeted Restoration of the Intestinal Microbiota with a Simple, Defined Bacteriotherapy Resolves Relapsing Clostridium difficile Disease in Mice , 2012, PLoS pathogens.

[7]  J. Tanha,et al.  Modulation of Toxin Production by the Flagellar Regulon in Clostridium difficile , 2012, Infection and Immunity.

[8]  C. Waters,et al.  A Tangled Web: Regulatory Connections between Quorum Sensing and Cyclic Di-GMP , 2012, Journal of bacteriology.

[9]  N. Fairweather,et al.  The Clostridium difficile spo0A Gene Is a Persistence and Transmission Factor , 2012, Infection and Immunity.

[10]  C. Waters,et al.  Cyclic Diguanylate Inversely Regulates Motility and Aggregation in Clostridium difficile , 2012, Journal of bacteriology.

[11]  A. Shen.  Clostridium difficile Toxins: Mediators of Inflammation , 2012, Journal of Innate Immunity.

[12]  K. Winzer,et al.  An agr Quorum Sensing System That Regulates Granulose Formation and Sporulation in Clostridium acetobutylicum , 2011, Applied and Environmental Microbiology.

[13]  Atina G. Coté,et al.  The draft genome and transcriptome of Cannabis sativa , 2011, Genome Biology.

[14]  G. Armstrong,et al.  Mutagenic Analysis of the Clostridium difficile Flagellar Proteins, FliC and FliD, and Their Contribution to Virulence in Hamsters , 2011, Infection and Immunity.

[15]  I. Martin-Verstraete,et al.  The Key Sigma Factor of Transition Phase, SigH, Controls Sporulation, Metabolism, and Virulence Factor Expression in Clostridium difficile , 2011, Journal of bacteriology.

[16]  V. Burrus,et al.  c-di-GMP Turn-Over in Clostridium difficile Is Controlled by a Plethora of Diguanylate Cyclases and Phosphodiesterases , 2011, PLoS genetics.

[17]  I. Martin-Verstraete,et al.  CcpA‐mediated repression of Clostridium difficile toxin gene expression , 2011, Molecular microbiology.

[18]  E. Papoutsakis,et al.  Small RNAs in the Genus Clostridium , 2011, mBio.

[19]  Nigel P. Minton,et al.  The role of toxin A and toxin B in Clostridium difficile infection , 2010, Nature.

[20]  J. Heap,et al.  The emergence of 'hypervirulence' in Clostridium difficile. , 2010, International journal of medical microbiology : IJMM.

[21]  M. Sebaihia,et al.  Array comparative hybridisation reveals a high degree of similarity between UK and European clinical isolates of hypervirulent Clostridium difficile , 2010, BMC Genomics.

[22]  Rodrigo Lopez,et al.  A new bioinformatics analysis tools framework at EMBL–EBI , 2010, Nucleic Acids Res..

[23]  Andries J. van Tonder,et al.  Evolutionary dynamics of Clostridium difficile over short and long time scales , 2010, Proceedings of the National Academy of Sciences.

[24]  A. Cheung,et al.  Faculty Opinions recommendation of Evolution of MRSA during hospital transmission and intercontinental spread. , 2010 .

[25]  M. Quail,et al.  Comparative genome and phenotypic analysis of Clostridium difficile 027 strains provides insight into the evolution of a hypervirulent bacterium , 2009, Genome Biology.

[26]  R. Lewis,et al.  Characterization of the Sporulation Initiation Pathway of Clostridium difficile and Its Role in Toxin Production , 2009, Journal of bacteriology.

[27]  J. Heap,et al.  A modular system for Clostridium shuttle plasmids. , 2009, Journal of microbiological methods.

[28]  N. Fairweather,et al.  Antibiotic Treatment of Clostridium difficile Carrier Mice Triggers a Supershedder State, Spore-Mediated Transmission, and Severe Disease in Immunocompromised Hosts , 2009, Infection and Immunity.

[29]  Julian I. Rood,et al.  Toxin B is essential for virulence of Clostridium difficile , 2009, Nature.

[30]  Jeffrey D Goldsmith,et al.  A mouse model of Clostridium difficile-associated disease. , 2008, Gastroenterology.

[31]  C. Kelly,et al.  Clostridium difficile--more difficult than ever. , 2008, The New England journal of medicine.

[32]  Holger Fröhlich,et al.  Predicting pathway membership via domain signatures , 2008, Bioinform..

[33]  R. Breaker,et al.  Riboswitches in Eubacteria Sense the Second Messenger Cyclic Di-GMP , 2008, Science.

[34]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[35]  D. Sidote,et al.  Structure of the Staphylococcus aureus AgrA LytTR domain bound to DNA reveals a beta fold with an unusual mode of binding. , 2008, Structure.

[36]  Rodrigo Lopez,et al.  Clustal W and Clustal X version 2.0 , 2007, Bioinform..

[37]  M. Babu,et al.  Conservation and Evolutionary Dynamics of the agr Cell-to-Cell Communication System across Firmicutes , 2007, Journal of bacteriology.

[38]  A. Sonenshein,et al.  Repression of Clostridium difficile toxin gene expression by CodY , 2007, Molecular microbiology.

[39]  L. Steinmetz,et al.  Antisense artifacts in transcriptome microarray experiments are resolved by actinomycin D , 2007, Nucleic acids research.

[40]  E. Kuijper,et al.  Spread and epidemiology of Clostridium difficile polymerase chain reaction ribotype 027/toxinotype III in The Netherlands. , 2007, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[41]  L. Mascola,et al.  Increase in Clostridium difficile–related Mortality Rates, United States, 1999–2004 , 2007, Emerging infectious diseases.

[42]  J. Heap,et al.  The ClosTron: a universal gene knock-out system for the genus Clostridium. , 2007, Journal of microbiological methods.

[43]  M. Wilcox,et al.  Clostridium difficile: changing epidemiology and new treatment options , 2007, Current opinion in infectious diseases.

[44]  M. Dionne,et al.  A portrait of the geographic dissemination of the Clostridium difficile North American pulsed-field type 1 strain and the epidemiology of C. difficile-associated disease in Québec. , 2007, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[45]  Julian Parkhill,et al.  The multidrug-resistant human pathogen Clostridium difficile has a highly mobile, mosaic genome , 2006, Nature Genetics.

[46]  S. Hilsenbeck,et al.  Comparison of OG1RF and an Isogenic fsrB Deletion Mutant by Transcriptional Analysis: the Fsr System of Enterococcus faecalis Is More than the Activator of Gelatinase and Serine Protease , 2006, Journal of bacteriology.

[47]  Ken Dewar,et al.  A predominantly clonal multi-institutional outbreak of Clostridium difficile-associated diarrhea with high morbidity and mortality. , 2005, The New England journal of medicine.

[48]  Stuart Johnson,et al.  An epidemic, toxin gene-variant strain of Clostridium difficile. , 2005, The New England journal of medicine.

[49]  K. Song,et al.  LuxS/autoinducer-2 quorum sensing molecule regulates transcriptional virulence gene expression in Clostridium difficile. , 2005, Biochemical and biophysical research communications.

[50]  Jon Brazier,et al.  Toxin production by an emerging strain of Clostridium difficile associated with outbreaks of severe disease in North America and Europe , 2005, The Lancet.

[51]  S. Maleki,et al.  Staphylococcus aureus AgrA Binding to the RNAIII-agr Regulatory Region , 2004, Journal of bacteriology.

[52]  C. Solano,et al.  Role of the GGDEF protein family in Salmonella cellulose biosynthesis and biofilm formation , 2004, Molecular microbiology.

[53]  U. Römling,et al.  GGDEF and EAL domains inversely regulate cyclic di‐GMP levels and transition from sessility to motility , 2004, Molecular microbiology.

[54]  F. Ausubel,et al.  Contribution of Gelatinase, Serine Protease, and fsr to the Pathogenesis of Enterococcus faecalis Endophthalmitis , 2004, Infection and Immunity.

[55]  A. Lambowitz,et al.  Use of computer-designed group II introns to disrupt Escherichia coli DExH/D-box protein and DNA helicase genes. , 2004, Journal of molecular biology.

[56]  A. Charbit,et al.  Identification of the agr Locus of Listeria monocytogenes: Role in Bacterial Virulence , 2003, Infection and Immunity.

[57]  A. McLeod,et al.  Conjugative transfer of clostridial shuttle vectors from Escherichia coli to Clostridium difficile through circumvention of the restriction barrier , 2002, Molecular microbiology.

[58]  Frederick M. Ausubel,et al.  Virulence Effect of Enterococcus faecalis Protease Genes and the Quorum-Sensing Locus fsr in Caenorhabditis elegans and Mice , 2002, Infection and Immunity.

[59]  F. Ausubel,et al.  The Enterococcus faecalis fsrB Gene, a Key Component of the fsr Quorum-Sensing System, Is Associated with Virulence in the Rabbit Endophthalmitis Model , 2002, Infection and Immunity.

[60]  H. Boureau,et al.  Role of FliC and FliD Flagellar Proteins ofClostridium difficile in Adherence and Gut Colonization , 2001, Infection and Immunity.

[61]  M. Rohde,et al.  The multicellular morphotypes of Salmonella typhimurium and Escherichia coli produce cellulose as the second component of the extracellular matrix , 2001, Molecular microbiology.

[62]  G. Weinstock,et al.  Effects of Enterococcus faecalis fsrGenes on Production of Gelatinase and a Serine Protease and Virulence , 2000, Infection and Immunity.

[63]  S. Arvidson,et al.  Regulation of agr-Dependent Virulence Genes in Staphylococcus aureus by RNAIII from Coagulase-Negative Staphylococci , 1998, Journal of bacteriology.

[64]  D. Gerding,et al.  Clostridium difficile--associated diarrhea. , 1998, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[65]  R. Beavis,et al.  Bacterial interference caused by autoinducing peptide variants. , 1997, Science.

[66]  J. Kornblum,et al.  Synthesis of staphylococcal virulence factors is controlled by a regulatory RNA molecule. , 1993, The EMBO journal.

[67]  M. Young,et al.  Conjugative plasmid transfer from Escherichia coli to Clostridium acetobutylicum. , 1990, Journal of general microbiology.

[68]  B. Wren,et al.  Restriction endonuclease DNA analysis of Clostridium difficile , 1987, Journal of clinical microbiology.

[69]  K. Wilson.  Efficiency of various bile salt preparations for stimulation of Clostridium difficile spore germination , 1983, Journal of clinical microbiology.

[70]  which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. MAnorm: a robust model for quantitative comparison of ChIP-Seq data sets , 2011 .

[71]  J. Heap,et al.  The ClosTron: Mutagenesis in Clostridium refined and streamlined. , 2010, Journal of microbiological methods.

[72]  M. O'Reilly,et al.  Regulation of exoprotein gene expression in Staphylococcus aureus by agr , 2004, Molecular and General Genetics MGG.

[73]  B. Tennison,et al.  Commission for Healthcare Audit and Inspection , 2004 .

[74]  Claude-Alain H. Roten,et al.  Theoretical and practical advances in genome halving , 2004 .