The RcsAB Box

The interaction of the two transcriptional regulators RcsA and RcsB with a specific operator is a common mechanism in the activation of capsule biosynthesis in enteric bacteria. We describe RcsAB binding sites in the wza promoter of the operon for colanic acid biosynthesis in Escherichia coliK-12, in the galF promoter of the operon for K2 antigen biosynthesis in Klebsiella pneumoniae, and in thetviA (vipR) promoter of the operon for Vi antigen biosynthesis in Salmonella typhi. We further show the interaction of RcsAB with the rcsA promoters of various species, indicating that rcsA autoregulation also depends on the presence of both proteins. The compilation of all identified RcsAB binding sites revealed the conserved core sequence TaAGaatatTCctA, which we propose to be termed RcsAB box. The RcsAB box is also part ofBordetella pertussis BvgA binding sites and may represent a more distributed recognition motif within the LuxR superfamily of transcriptional regulators. The RcsAB box is essential for the induction of Rcs-regulated promoters. Site-specific mutations of conserved nucleotides in the RcsAB boxes of the E. coli wzaand rcsA promoters resulted in an exopolysaccharide-negative phenotype and in the reduction of reporter gene expression.

[1]  J. O. Lee,et al.  Characterization of the rcsA and rcsB genes from Salmonella typhi: rcsB through tviA is involved in regulation of Vi antigen synthesis , 1996, Journal of bacteriology.

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

[3]  M. Coleman,et al.  Molecular cloning, expression and nucleotide sequence of the rcsA gene of Erwinia amylovora, encoding a positive regulator of capsule expression: evidence for a family of related capsule activator proteins. , 1990, Journal of general microbiology.

[4]  K. Geider,et al.  The rcsA gene from Erwinia amylovora: identification, nucleotide sequence, and regulation of exopolysaccharide biosynthesis. , 1990, Molecular plant-microbe interactions : MPMI.

[5]  C. Locht,et al.  The modular architecture of bacterial response regulators. Insights into the activation mechanism of the BvgA transactivator of Bordetella pertussis. , 1994, Journal of molecular biology.

[6]  R. Rappuoli,et al.  Positive transcriptional feedback at the bvg locus controls expression of virulence factors in Bordetella pertussis. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[7]  S. Gottesman,et al.  RcsA, an unstable positive regulator of capsular polysaccharide synthesis , 1991, Journal of bacteriology.

[8]  F. Smith,et al.  COLORIMETRIC METHOD FOR DETER-MINATION OF SUGAR AND RELATED SUBSTANCE , 1956 .

[9]  S. Gottesman,et al.  Fine-structure mapping and identification of two regulators of capsule synthesis in Escherichia coli K-12 , 1988, Journal of bacteriology.

[10]  S. Falkow,et al.  Identification of Bordetella pertussis regulatory sequences required for transcriptional activation of the fhaB gene and autoregulation of the bvgAS operon , 1991, Journal of bacteriology.

[11]  C. Whitfield,et al.  expression of group I capsular of the role of the rcs regulatory system in Escherichia coli O 9 : K 30 : H 12 and examination Characterization of rcsB and rcsC from , 2022 .

[12]  K. Geider,et al.  Characterization of the rcsB gene from Erwinia amylovora and its influence on exoploysaccharide synthesis and virulence of the fire blight pathogen , 1997, Journal of bacteriology.

[13]  S. Gottesman,et al.  Capsule synthesis in Escherichia coli K-12 is regulated by proteolysis , 1987, Journal of bacteriology.

[14]  J. Saunders,et al.  Isolation from Klebsiella and characterization of two rcs genes that activate colanic acid capsular biosynthesis in Escherichia coli. , 1987, Journal of general microbiology.

[15]  S. Gottesman,et al.  Regulation of capsular polysaccharide synthesis in Escherichia coli K-12: characterization of three regulatory genes , 1985, Journal of bacteriology.

[16]  S. Gottesman,et al.  Regulation of capsular polysaccharide synthesis in Escherichia coli K12 , 1991, Molecular microbiology.

[17]  T. Ezaki,et al.  Complete nucleotide sequence and molecular characterization of ViaB region encoding Vi antigen in Salmonella typhi , 1993, Journal of bacteriology.

[18]  W. Saenger,et al.  Identification of an RcsA/RcsB Recognition Motif in the Promoters of Exopolysaccharide Biosynthetic Operons from Erwinia amylovora and Pantoea stewartii Subspeciesstewartii * , 1999, The Journal of Biological Chemistry.

[19]  R. Rappuoli,et al.  Differential binding of BvgA to two classes of virulence genes of Bordetella pertussis directs promoter selectivity by RNA polymerase , 1996, Molecular microbiology.

[20]  E. E. Baker,et al.  The VI antigens of the Enterobacteriaceae. I. Purification and chemical properties. , 1959, Journal of immunology.

[21]  J. Leigh,et al.  Exopolysaccharides in plant-bacterial interactions. , 1992, Annual review of microbiology.

[22]  S. Gottesman,et al.  RcsB and RcsC: a two-component regulator of capsule synthesis in Escherichia coli , 1990, Journal of bacteriology.

[23]  J. Trempy,et al.  Identification of a second RcsA protein, a positive regulator of colanic acid capsular polysaccharide genes, in Escherichia coli , 1996, Journal of bacteriology.

[24]  A. Ishihama,et al.  Protein-protein communication within the transcription apparatus , 1993, Journal of bacteriology.

[25]  A. Chatterjee,et al.  Isolation and characterization of an rcsA-like gene of Erwinia amylovora that activates extracellular polysaccharide production in Erwinia species, Escherichia coli, and Salmonella typhimurium , 1990 .

[26]  V. Stout.  Identification of the promoter region for the colanic acid polysaccharide biosynthetic genes in Escherichia coli K-12 , 1996, Journal of bacteriology.

[27]  K. Geider,et al.  Interaction of the regulator proteins RcsA and RcsB with the promoter of the operon for amylovoran biosynthesis in Erwinia amylovora , 1997, Molecular and General Genetics MGG.

[28]  E. Greenberg,et al.  Synergistic binding of the Vibrio fischeri LuxR transcriptional activator domain and RNA polymerase to the lux promoter region , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[29]  K. Murakami,et al.  Nature of DNA binding and RNA polymerase interaction of the Bordetella pertussis BvgA transcriptional activator at the fha promoter , 1997, Journal of bacteriology.

[30]  T. Ezaki,et al.  Positive autoregulation of vipR expression in ViaB region-encoded Vi antigen of Salmonella typhi , 1996, Journal of bacteriology.

[31]  T. Baldwin,et al.  Identification of the operator of the lux regulon from the Vibrio fischeri strain ATCC7744. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[32]  S. Gottesman,et al.  Osmotic shock induction of capsule synthesis in Escherichia coli K-12 , 1996, Journal of bacteriology.

[33]  P. Reeves,et al.  Organization of the Escherichia coli K-12 gene cluster responsible for production of the extracellular polysaccharide colanic acid , 1996, Journal of bacteriology.

[34]  S. Henikoff,et al.  Finding protein similarities with nucleotide sequence databases. , 1990, Methods in enzymology.

[35]  J. Trempy,et al.  Escherichia coli RcsA, a Positive Activator of Colanic Acid Capsular Polysaccharide Synthesis, Functions To Activate Its Own Expression , 1999, Journal of bacteriology.

[36]  S. Stibitz.  Mutations in the bvgA gene of Bordetella pertussis that differentially affect regulation of virulence determinants , 1994, Journal of bacteriology.

[37]  L. Baron,et al.  Expression of Vi antigen in Escherichia coli K-12: characterization of ViaB from Citrobacter freundii and identity of ViaA with RcsB , 1992, Journal of bacteriology.

[38]  C. Whitfield,et al.  The rcsA gene of Klebsiella pneumoniae O1:K20 is involved in expression of the serotype-specific K (capsular) antigen , 1991, Infection and immunity.

[39]  P. Bugert,et al.  Molecular analysis of the ams operon required for exopolysaccharide synthesis of Erwinia amylovora , 1995, Molecular microbiology.

[40]  R. Pitt,et al.  A NEW ANTIGEN OF B. TYPHOSUS: ITS RELATION TO VIRULENCE AND TO ACTIVE AND PASSIVE IMMUNISATION , 1934 .

[41]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[42]  Michael Gribskov,et al.  Combining evidence using p-values: application to sequence homology searches , 1998, Bioinform..

[43]  Y. Arakawa,et al.  Genomic organization of the Klebsiella pneumoniae cps region responsible for serotype K2 capsular polysaccharide synthesis in the virulent strain Chedid , 1995, Journal of bacteriology.

[44]  N. W. Davis,et al.  The complete genome sequence of Escherichia coli K-12. , 1997, Science.

[45]  K. Geider,et al.  A plasmid cloning system utilizing replication and packaging functions of the filamentous bacteriophage fd. , 1985, Gene.