P-Ser-HPr--a link between carbon metabolism and the virulence of some pathogenic bacteria.

HPr kinase/phosphorylase phosphorylates HPr, a phosphocarrier protein of the phosphoenolpyruvate:carbohydrate phosphotransferase system, at serine-46. P-Ser-HPr is the central regulator of carbon metabolism in Gram-positive bacteria, but also plays a role in virulence development of certain pathogens. In Listeria monocytogenes, several virulence genes, which depend on the transcription activator PrfA, are repressed by glucose, fructose, etc., in a catabolite repressor (CcpA)-independent mechanism. However, the catabolite co-repressor P-Ser-HPr was found to inhibit the activity of PrfA. In an hprKV267F mutant, in which most of the HPr is transformed into P-Ser-HPr, PrfA was barely active. The ptsH1 mutation (Ser-46 of HPr replaced with an alanine) prevented the inhibitory effect of the hprKV267F mutation. Interestingly, disruption of ccpA also inhibited PrfA activity. This effect is probably also mediated via P-Ser-HPr, since ccpA disruption leads to elevated amounts of P-Ser-HPr. Indeed, a ccpA ptsH1 double mutant exhibited normal PrfA activity. In S. pyogenes, the expression of several virulence genes depends on the transcription activator Mga. Interestingly, the mga promoter is preceded by an operator site, which serves as target for the CcpA/P-Ser-HPr complex. Numerous Gram-negative pathogens also contain hprK, which is often organised in an operon with transcription regulators necessary for the development of virulence, indicating that in these organisms P-Ser-HPr also plays a role in pathogenesis. Indeed, inactivation of Neisseria meningitidis hprK strongly diminished cell adhesion of this pathogen.

[1]  G. Nimmo,et al.  Partial purification and properties of isocitrate dehydrogenase kinase/phosphatase from Escherichia coli ML308. , 1984, European journal of biochemistry.

[2]  J. W. Neal,et al.  Regulation of the glucose:H+ symporter by metabolite-activated ATP-dependent phosphorylation of HPr in Lactobacillus brevis , 1994, Journal of bacteriology.

[3]  S. Nessler,et al.  The bacterial HPr kinase/phosphorylase: a new type of Ser/Thr kinase as antimicrobial target. , 2005, Biochimica et biophysica acta.

[4]  J. Deutscher,et al.  The hprK gene of Enterococcus faecalis encodes a novel bifunctional enzyme: the HPr kinase/phosphatase , 1999, Molecular microbiology.

[5]  Christophe Geourjon,et al.  A New Family of Phosphotransferases with a P-loop Motif* , 2002, The Journal of Biological Chemistry.

[6]  S. Cole,et al.  Proteomic identification of M. tuberculosis protein kinase substrates: PknB recruits GarA, a FHA domain-containing protein, through activation loop-mediated interactions. , 2005, Journal of molecular biology.

[7]  Trevor C. Charles,et al.  A chromosomally encoded two-component sensory transduction system is required for virulence of Agrobacterium tumefaciens , 1993, Journal of bacteriology.

[8]  M. Pallen,et al.  Bacterial FHA domains: neglected players in the phospho-threonine signalling game? , 2002, Trends in microbiology.

[9]  K. E. Pullen,et al.  An alternate conformation and a third metal in PstP/Ppp, the M. tuberculosis PP2C-Family Ser/Thr protein phosphatase. , 2004, Structure.

[10]  P. Glaser,et al.  Salt Stress Proteins Induced in Listeria monocytogenes , 2002, Applied and Environmental Microbiology.

[11]  W. Carmichael,et al.  Cyanobacterial PPP Family Protein Phosphatases Possess Multifunctional Capabilities and Are Resistant to Microcystin-LR* , 1999, The Journal of Biological Chemistry.

[12]  R. Losick,et al.  Bacillus Subtilis and Its Closest Relatives: From Genes to Cells , 2001 .

[13]  First structural glimpse at a bacterial Ser/Thr protein phosphatase. , 2004, Structure.

[14]  A. Cozzone,et al.  Characterization of the phosphoproteins of Escherichia coli cells by electrophoretic analysis. , 1986, European journal of biochemistry.

[15]  V. Monedero,et al.  Phosphorylation of HPr by the Bifunctional HPr Kinase/P-Ser-HPr Phosphatase from Lactobacillus casei Controls Catabolite Repression and Inducer Exclusion but Not Inducer Expulsion , 2000, Journal of bacteriology.

[16]  S. Raina,et al.  Phosphorylation‐mediated regulation of heat shock response in Escherichia coli , 2003, Molecular microbiology.

[17]  A. Cozzone,et al.  Autophosphorylation of a bacterial protein at tyrosine. , 1996, Journal of molecular biology.

[18]  S. Séror,et al.  PrpE, a PPP protein phosphatase from Bacillus subtilis with unusual substrate specificity. , 2002, The Biochemical journal.

[19]  P. Youngman,et al.  Carbon‐source regulation of virulence gene expression in Listeria monocytogenes , 1997, Molecular microbiology.

[20]  P. Youngman,et al.  A Homolog of CcpA Mediates Catabolite Control in Listeria monocytogenes but Not Carbon Source Regulation of Virulence Genes , 1998 .

[21]  B. Kreikemeyer,et al.  Virulence factor regulation and regulatory networks in Streptococcus pyogenes and their impact on pathogen-host interactions. , 2003, Trends in microbiology.

[22]  J. Deutscher,et al.  Transcription Regulators Potentially Controlled by HPr Kinase/Phosphorylase in Gram-Negative Bacteria , 2003, Journal of Molecular Microbiology and Biotechnology.

[23]  V. Molle,et al.  Two FHA domains on an ABC transporter, Rv1747, mediate its phosphorylation by PknF, a Ser/Thr protein kinase from Mycobacterium tuberculosis. , 2004, FEMS microbiology letters.

[24]  Wolfgang Hengstenberg,et al.  Structure of the full-length HPr kinase/phosphatase from Staphylococcus xylosus at 1.95 Å resolution: Mimicking the product/substrate of the phospho transfer reactions , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[25]  K. Kobayashi,et al.  Combined transcriptome and proteome analysis as a powerful approach to study genes under glucose repression in Bacillus subtilis. , 2001, Nucleic acids research.

[26]  J. Yother,et al.  CpsB Is a Modulator of Capsule-associated Tyrosine Kinase Activity in Streptococcus pneumoniae * , 2001, The Journal of Biological Chemistry.

[27]  D. Laporte,et al.  A single gene codes for the kinase and phosphatase which regulate isocitrate dehydrogenase. , 1985, The Journal of biological chemistry.

[28]  H. Reeves,et al.  Phosphorylation of Isocitrate dehydrogenase of Escherichia coli. , 1979, Science.

[29]  Mutational Analysis of the Role of HPr inListeria monocytogenes , 1999, Applied and Environmental Microbiology.

[30]  D. Petranovic,et al.  In Vitro Characterization of the Bacillus subtilis Protein Tyrosine Phosphatase YwqE , 2005, Journal of bacteriology.

[31]  M. Hecker,et al.  Transcriptional analysis of bglPH expression in Bacillus subtilis: evidence for two distinct pathways mediating carbon catabolite repression , 1996, Journal of bacteriology.

[32]  J. Deutscher,et al.  Autophosphorylation of the Escherichia coli Protein Kinase Wzc Regulates Tyrosine Phosphorylation of Ugd, a UDP-glucose Dehydrogenase* , 2003, Journal of Biological Chemistry.

[33]  R. Losick,et al.  Activation of Cell-Specific Transcription by a Serine Phosphatase at the Site of Asymmetric Division , 1995, Science.

[34]  P. Cossart,et al.  Differential activation of virulence gene expression by PrfA, the Listeria monocytogenes virulence regulator , 1995, Journal of bacteriology.

[35]  V. Monedero,et al.  Mutations lowering the phosphatase activity of HPr kinase/phosphatase switch off carbon metabolism , 2001, The EMBO journal.

[36]  K. McIver,et al.  Transcriptional Activation of sclA by Mga Requires a Distal Binding Site in Streptococcus pyogenes , 2004, Journal of bacteriology.

[37]  B. Poolman,et al.  Phosphorylation State of HPr Determines the Level of Expression and the Extent of Phosphorylation of the Lactose Transport Protein ofStreptococcus thermophilus * , 2000, The Journal of Biological Chemistry.

[38]  J. Janin,et al.  X‐ray structure of HPr kinase: a bacterial protein kinase with a P‐loop nucleotide‐binding domain , 2001, The EMBO journal.

[39]  Y. Auffray,et al.  Characterization of the ccpA Gene ofEnterococcus faecalis: Identification of Starvation-Inducible Proteins Regulated by CcpA , 2000, Journal of bacteriology.

[40]  J. Deutscher,et al.  Transmembrane modulator‐dependent bacterial tyrosine kinase activates UDP‐glucose dehydrogenases , 2003, The EMBO journal.

[41]  W. Weyler,et al.  Catabolite repression mediated by the CcpA protein in Bacillus subtilis: novel modes of regulation revealed by whole‐genome analyses , 2001, Molecular microbiology.

[42]  E. Sutherland,et al.  Inactivation and Activation of Liver Phosphorylase , 1955, Nature.

[43]  M. Saier,et al.  ATP-dependent protein kinase-catalyzed phosphorylation of a seryl residue in HPr, a phosphate carrier protein of the phosphotransferase system in Streptococcus pyogenes. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Tom Alber,et al.  Structure of Mycobacterium tuberculosis PknB supports a universal activation mechanism for Ser/Thr protein kinases , 2003, Nature Structural Biology.

[45]  J. Lowy,et al.  Roles of pilin and PilC in adhesion of Neisseria meningitidis to human epithelial and endothelial cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[46]  S. Séror,et al.  Characterization of PrpC from Bacillus subtilis, a Member of the PPM Phosphatase Family , 2000, Journal of bacteriology.

[47]  J. Deutscher,et al.  New protein kinase and protein phosphatase families mediate signal transduction in bacterial catabolite repression. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[48]  R. Brennan,et al.  Crystal structure of HPr kinase/phosphatase from Mycoplasma pneumoniae. , 2003, Journal of molecular biology.

[49]  O. Kuipers,et al.  Regulatory Functions of Serine-46-Phosphorylated HPr in Lactococcus lactis , 2001, Journal of bacteriology.

[50]  J. Deutscher,et al.  Purification and characterization of an ATP-dependent protein kinase from Streptococcus faecalis , 1984 .

[51]  June R. Scott,et al.  Regulation of mga Transcription in the Group A Streptococcus: Specific Binding of Mga within Its Own Promoter and Evidence for a Negative Regulator , 1999, Journal of bacteriology.

[52]  P. Alzari,et al.  Crystal Structure of the Catalytic Domain of the PknB Serine/Threonine Kinase from Mycobacterium tuberculosis * , 2003, The Journal of Biological Chemistry.

[53]  Ivan Mijakovic,et al.  X-ray structure of a bifunctional protein kinase in complex with its protein substrate HPr , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[54]  Wolf-Dieter Schubert,et al.  The mutation G145S in PrfA, a key virulence regulator of Listeria monocytogenes, increases DNA‐binding affinity by stabilizing the HTH motif , 2005, Molecular microbiology.

[55]  J. Stülke,et al.  Control of the glycolytic gapA operon by the catabolite control protein A in Bacillus subtilis: a novel mechanism of CcpA‐mediated regulation , 2002, Molecular microbiology.

[56]  Ivan Mijakovic,et al.  Pyrophosphate-producing protein dephosphorylation by HPr kinase/phosphorylase: A relic of early life? , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[57]  Isabelle Martin-Verstraete,et al.  Carbohydrate Uptake and Metabolism , 2002 .

[58]  J. Deutscher,et al.  The Bacillus subtilis crh gene encodes a HPr-like protein involved in carbon catabolite repression. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[59]  J. Deutscher,et al.  Antitermination by GlpP, catabolite repression via CcpA and inducer exclusion triggered by P~GlpK dephosphorylation control Bacillus subtilis glpFK expression , 2002, Molecular microbiology.

[60]  Y. Fujita,et al.  Specific recognition of the Bacillus subtilis gnt cis‐acting catabolite‐responsive element by a protein complex formed between CcpA and seryl‐phosphorylated HPr , 1995, Molecular microbiology.

[61]  D. Durocher,et al.  The molecular basis of FHA domain:phosphopeptide binding specificity and implications for phospho-dependent signaling mechanisms. , 2000, Molecular cell.

[62]  G. Rapoport,et al.  Antagonistic effects of dual PTS‐catalysed phosphorylation on the Bacillus subtilis transcriptional activator LevR , 1998, Molecular microbiology.

[63]  S. Inouye,et al.  A gene encoding a protein serine/threonine kinase is required for normal development of M. xanthus, a gram-negative bacterium , 1991, Cell.

[64]  E. Krebs,et al.  Conversion of phosphorylase b to phosphorylase a in muscle extracts. , 1955, The Journal of biological chemistry.

[65]  W. Hillen,et al.  Protein kinase‐dependent HPr/CcpA interaction links glycolytic activity to carbon catabolite repression in Gram‐positive bacteria , 1995, Molecular microbiology.

[66]  J. R. Scott,et al.  Role of mga in growth phase regulation of virulence genes of the group A streptococcus , 1997, Journal of bacteriology.

[67]  P. Postma,et al.  Phosphoenolpyruvate:carbohydrate phosphotransferase system of bacteria. , 1985, Microbiological reviews.

[68]  M. Schumacher,et al.  Structural Basis for Allosteric Control of the Transcription Regulator CcpA by the Phosphoprotein HPr-Ser46-P , 2004, Cell.