SlyA and H-NS Regulate Transcription of the Escherichia coli K5 Capsule Gene Cluster, and Expression of slyA in Escherichia coli Is Temperature-dependent, Positively Autoregulated, and Independent of H-NS*

In this paper, we present the first evidence of a role for the transcriptional regulator SlyA in the regulation of transcription of the Escherichia coli K5 capsule gene cluster and demonstrate, using a combination of reporter gene fusions, DNase I footprinting, and electrophoretic mobility shift assays, the dependence of transcription on the functional interplay between H-NS and SlyA. Both SlyA and H-NS bind to multiple overlapping sites within the promoter in vitro, but their binding is not mutually exclusive, resulting in a remodeled nucleoprotein complex. In addition, we show that expression of the E. coli slyA gene is temperature-regulated, positively autoregulated, and independent of H-NS.

[1]  W. Goebel,et al.  Analysis of the SlyA‐controlled expression, subcellular localization and pore‐forming activity of a 34 kDa haemolysin (ClyA) from Escherichia coli K‐12 , 1999, Molecular microbiology.

[2]  B. Beatrix,et al.  Analysis of RovA, a Transcriptional Regulator of Yersinia pseudotuberculosis Virulence That Acts through Antirepression and Direct Transcriptional Activation* , 2005, Journal of Biological Chemistry.

[3]  C. Dorman,et al.  Domain organization and oligomerization among H-NS-like nucleoid-associated proteins in bacteria. , 1999, Trends in microbiology.

[4]  B. Kempf,et al.  Interactions of the nucleoid-associated DNA-binding protein H-NS with the regulatory region of the osmotically controlled proU operon of Escherichia coli. , 1994, The Journal of biological chemistry.

[5]  S. Crosby,et al.  RovA, a global regulator of Yersinia pestis, specifically required for bubonic plague , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[6]  D. Ussery,et al.  The chromatin‐associated protein H‐NS alters DNA topology in vitro. , 1994, The EMBO journal.

[7]  B. Uhlin,et al.  H-NS and StpA Proteins Stimulate Expression of the Maltose Regulon in Escherichia coli , 1998, Journal of bacteriology.

[8]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .

[9]  A. Flower,et al.  BipA is required for growth of Escherichia coli K12 at low temperature , 2001, Molecular Genetics and Genomics.

[10]  F. Fang,et al.  Co‐regulation of Salmonella enterica genes required for virulence and resistance to antimicrobial peptides by SlyA and PhoP/PhoQ , 2005, Molecular microbiology.

[11]  I. Roberts,et al.  Regulation of Escherichia coli K5 capsular polysaccharide expression: evidence for involvement of RfaH in the expression of group II capsules. , 1994, FEMS microbiology letters.

[12]  Antoine Danchin,et al.  Large‐scale monitoring of pleiotropic regulation of gene expression by the prokaryotic nucleoid‐associated protein, H‐NS , 2001, Molecular microbiology.

[13]  I. Roberts,et al.  Regulation of the Escherichia coli K5 capsule gene cluster by transcription antitermination , 1997, Molecular microbiology.

[14]  R. Wagner,et al.  LRP and H‐NS – cooperative partners for transcription regulation at Escherichia coli rRNA promoters , 2005, Molecular microbiology.

[15]  M. Woodward,et al.  Co‐ordination of pathogenicity island expression by the BipA GTPase in enteropathogenic Escherichia coli (EPEC) , 2003, Molecular microbiology.

[16]  C. Whitfield,et al.  Transcriptional organization and regulation of the Escherichia coli K30 group 1 capsule biosynthesis (cps) gene cluster , 2003, Molecular microbiology.

[17]  P. Reeves,et al.  The JUMPstart sequence: a 39 bp element common to several polysaccharide gene clusters , 1994, Molecular microbiology.

[18]  Yipeng Wang,et al.  Selective Silencing of Foreign DNA with Low GC Content by the H-NS Protein in Salmonella , 2006, Science.

[19]  Enhanced Binding of Altered H-NS Protein to Flagellar Rotor Protein FliG Causes Increased Flagellar Rotational Speed and Hypermotility in Escherichia coli* , 1998, The Journal of Biological Chemistry.

[20]  D. Žgur-Bertok,et al.  H-NS and Lrp serve as positive modulators of traJ expression from the Escherichia coli plasmid pRK100 , 2003, Molecular Genetics and Genomics.

[21]  P. Dersch,et al.  Environmental control of invasin expression in Yersinia pseudotuberculosis is mediated by regulation of RovA, a transcriptional activator of the SlyA/Hor family , 2001, Molecular microbiology.

[22]  Jeffrey Green,et al.  Regulation of Escherichia coli Hemolysin E Expression by H-NS and Salmonella SlyA , 2004, Journal of bacteriology.

[23]  I. Roberts,et al.  Regulation of the Escherichia coli K5 Capsule Gene Cluster: Evidence for the Roles of H-NS, BipA, and Integration Host Factor in Regulation of Group 2 Capsule Gene Clusters in Pathogenic E. coli , 2000, Journal of bacteriology.

[24]  I S Roberts,et al.  Structure, assembly and regulation of expression of capsules in Escherichia coli , 1999, Molecular microbiology.

[25]  H. Buc,et al.  Probing the structure, function, and interactions of the Escherichia coli H-NS and StpA proteins by using dominant negative derivatives , 1996, Journal of bacteriology.

[26]  H. Mollenkopf,et al.  SlyA, a regulatory protein from Salmonella typhimurium, induces a haemolytic and pore-forming protein in Escherichia coli , 1995, Molecular and General Genetics MGG.

[27]  I. Roberts,et al.  Transcriptional organization and regulation of expression of region 1 of the Escherichia coli K5 capsule gene cluster , 1996, Journal of bacteriology.

[28]  B. Uhlin,et al.  Silencing and Activation of ClyA Cytotoxin Expression in Escherichia coli , 2000, Journal of bacteriology.

[29]  C. Dorman.  H-NS: a universal regulator for a dynamic genome , 2004, Nature Reviews Microbiology.

[30]  A. C. Chang,et al.  Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid , 1978, Journal of bacteriology.

[31]  R. Wagner,et al.  Conformational changes of the upstream DNA mediated by H-NS and FIS regulate E. coli RrnB P1 promoter activity. , 1999, Journal of molecular biology.

[32]  R. Wagner,et al.  The Bacterial Regulatory Protein H-NS A Versatile Modulator of Nucleic Acid Structures , 2002, Biological chemistry.

[33]  P. Revell,et al.  A chromosomally encoded regulator is required for expression of the Yersinia enterocolitica inv gene and for virulence , 2000, Molecular microbiology.

[34]  W. Goebel,et al.  Differential Regulation of Multiple Proteins of Escherichia coli and Salmonella enterica Serovar Typhimurium by the Transcriptional Regulator SlyA , 2002, Journal of bacteriology.

[35]  F. Fang,et al.  SlyA, a transcriptional regulator of Salmonella typhimurium, is required for resistance to oxidative stress and is expressed in the intracellular environment of macrophages , 1997, Infection and immunity.

[36]  H. Buc,et al.  The Degree of Oligomerization of the H-NS Nucleoid Structuring Protein Is Related to Specific Binding to DNA* , 2002, The Journal of Biological Chemistry.

[37]  V. L. Miller,et al.  Regulation of virulence by members of the MarR/SlyA family. , 2006, Current opinion in microbiology.

[38]  V. L. Miller,et al.  H-NS Represses inv Transcription in Yersinia enterocolitica through Competition with RovA and Interaction with YmoA , 2006, Journal of bacteriology.

[39]  R. Read,et al.  Interaction of the Salmonella typhimuriumTranscription and Virulence Factor SlyA with Target DNA and Identification of Members of the SlyA Regulon* , 2002, The Journal of Biological Chemistry.

[40]  P. Dersch,et al.  RovA is autoregulated and antagonizes H‐NS‐mediated silencing of invasin and rovA expression in Yersinia pseudotuberculosis , 2004, Molecular microbiology.

[41]  W. Goebel,et al.  A cytolysin encoded by Salmonella is required for survival within macrophages. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[42]  R. Simons,et al.  Improved single and multicopy lac-based cloning vectors for protein and operon fusions. , 1987, Gene.

[43]  C. Whitfield.  Biosynthesis and assembly of capsular polysaccharides in Escherichia coli. , 2006, Annual review of biochemistry.

[44]  Eduardo A. Groisman,et al.  Transcriptional Control of the Antimicrobial Peptide Resistance ugtL Gene by the Salmonella PhoP and SlyA Regulatory Proteins* , 2004, Journal of Biological Chemistry.

[45]  I. Roberts,et al.  The cell surface expression of group 2 capsular polysaccharides in Escherichia coli: the role of KpsD, RhsA and a multi‐protein complex at the pole of the cell , 2006, Molecular microbiology.

[46]  W. Gilbert,et al.  Sequencing end-labeled DNA with base-specific chemical cleavages. , 1980, Methods in enzymology.

[47]  K. Tilly,et al.  Temperature-regulated expression of bacterial virulence genes. , 2000, Microbes and infection.

[48]  Paul Skipp,et al.  A dedicated translation factor controls the synthesis of the global regulator Fis , 2004, The EMBO journal.

[49]  B. Colonna,et al.  Involvement of FIS in the H‐NS‐mediated regulation of virF gene of Shigella and enteroinvasive Escherichia coli  ‡ , 2001, Molecular microbiology.