Selective Regulation of ptsG Expression by Fis

Transcription of ptsG encoding glucose-specific permease, enzyme IICBGlc, inEscherichia coli is initiated from two promoters, P1 and P2. ptsG transcription is repressed by Mlc, a glucose-inducible regulator of carbohydrate metabolism. The regulation of ptsG P1 transcription is also under positive control by cyclic AMP receptor protein and cyclic AMP complex (CRP·cAMP) as observed in other Mlc regulon. We report here that Fis, one of the nucleoid-associated proteins, plays a key role in glucose induction of Mlc regulon. ptsG transcription was induced when wild-type cells were grown in the presence of glucose. However, in afis mutant, the basal level of ptsGtranscription was higher but decreased when cells were grown in the presence of glucose, which implies the possibility of regulatory interactions among Fis, Mlc, and CRP·cAMP. Footprinting experiments with various probes and transcription assays revealed that Fis assists both Mlc repression and CRP·cAMP activation of ptsG P1 through the formation of Fis·CRP·Mlc or Fis·CRP nucleoprotein complexes at ptsG P1 promoter depending on the availability of glucose in the growth medium. ptsG P2 transcription was inhibited by Fis and Mlc. Tighter Mlc repression and enhanced CRP·cAMP activation of ptsG P1 by Fis enable cells to regulate Mlc regulon efficiently by selectively controlling the concentration of enzyme IICBGlc that modulates Mlc activity.

[1]  W. Boos,et al.  Negative transcriptional regulation of a positive regulator: the expression of malT, encoding the transcriptional activator of the maltose regulon of Escherichia coli, is negatively controlled by Mlc , 1998, Molecular microbiology.

[2]  C. Ball,et al.  Dramatic changes in Fis levels upon nutrient upshift in Escherichia coli , 1992, Journal of bacteriology.

[3]  H. Aiba,et al.  A novel regulatory role of glucose transporter of Escherichia coli: membrane sequestration of a global repressor Mlc , 2000, The EMBO journal.

[4]  H. Aiba,et al.  A lowered concentration of cAMP receptor protein caused by glucose is an important determinant for catabolite repression in Escherichia coli , 1993, Molecular microbiology.

[5]  H. Kornberg,et al.  Inducible phosphoenolpyruvate-dependent hexose phosphotransferase activities in Escherichia coli. , 1972, The Biochemical journal.

[6]  M. Saier,et al.  Regulation of carbohydrate uptake and adenylate cyclase activity mediated by the enzymes II of the phosphoenolpyruvate: sugar phosphotransferase system in Escherichia coli. , 1976, The Journal of biological chemistry.

[7]  F. Neidhardt,et al.  Escherichia Coli and Salmonella: Typhimurium Cellular and Molecular Biology , 1987 .

[8]  S. Ryu,et al.  Purification of Mlc and Analysis of Its Effects on thepts Expression in Escherichia coli * , 1999, The Journal of Biological Chemistry.

[9]  S. Altuvia,et al.  Escherichia coli response to hydrogen peroxide: a role for DNA supercoiling, Topoisomerase I and Fis , 2000, Molecular microbiology.

[10]  H. Buc,et al.  On the action of the cyclic AMP‐cyclic AMP receptor protein complex at the Escherichia coli lactose and galactose promoter regions. , 1984, The EMBO journal.

[11]  J. Plumbridge.  Control of the expression of the manXYZ operon in Escherichia coli: Mlc is a negative regulator of the mannose PTS , 1998, Molecular microbiology.

[12]  J. St. Geme, III,et al.  Influence of pili, fibrils, and capsule on in vitro adherence by Haemophilus influenzae type b , 1996, Molecular microbiology.

[13]  Dongwoo Shin,et al.  The Escherichia coli glucose transporter enzyme IICBGlc recruits the global repressor Mlc , 2001, The EMBO journal.

[14]  S. Garges,et al.  Promoter switch in the Escherichia coli pts operon. , 1994, The Journal of biological chemistry.

[15]  W. Boos,et al.  Signal transduction between a membrane‐bound transporter, PtsG, and a soluble transcription factor, Mlc, of Escherichia coli , 2000, The EMBO journal.

[16]  B. Erni,et al.  The glucose transporter of Escherichia coli. Overexpression, purification, and characterization of functional domains. , 1994, The Journal of biological chemistry.

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

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

[19]  R. Ebright,et al.  Transcription activation by catabolite activator protein (CAP). , 1999, Journal of molecular biology.

[20]  S. McLeod,et al.  Coactivation of the RpoS-Dependent proPP2 Promoter by Fis and Cyclic AMP Receptor Protein , 2000, Journal of bacteriology.

[21]  J. B. McNeil,et al.  A simple in vivo footprinting method to examine DNA-protein interactions over the yeast PYK UAS element. , 1994, Nucleic acids research.

[22]  R. Ebright,et al.  Transcription activation at Class II CAP‐dependent promoters , 1993, Molecular microbiology.

[23]  T. Inada,et al.  A global repressor (Mlc) is involved in glucose induction of the ptsG gene encoding major glucose transporter in Escherichia coli , 1998, Molecular microbiology.

[24]  A. Travers,et al.  CRP Modulates fis Transcription by Alternate Formation of Activating and Repressing Nucleoprotein Complexes* , 2001, The Journal of Biological Chemistry.

[25]  P. Postma,et al.  cAMP receptor protein-cAMP plays a crucial role in glucose-lactose diauxie by activating the major glucose transporter gene in Escherichia coli. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[26]  S. Ryu,et al.  Heat Shock RNA Polymerase (Eς32) Is Involved in the Transcription of mlc and Crucial for Induction of the Mlc Regulon by Glucose in Escherichia coli * , 2001, The Journal of Biological Chemistry.

[27]  R Kahmann,et al.  Regulation of crp transcription by oscillation between distinct nucleoprotein complexes , 1998, The EMBO journal.

[28]  S. Ueda,et al.  Growth Phase-Dependent Variation in Protein Composition of the Escherichia coli Nucleoid , 1999, Journal of bacteriology.

[29]  T. D. Schneider,et al.  Information analysis of Fis binding sites. , 1997, Nucleic acids research.

[30]  T. Inada,et al.  Negative regulation of the pts operon by Mlc: mechanism underlying glucose induction in Escherichia coli , 1999, Genes to cells : devoted to molecular & cellular mechanisms.

[31]  T. Inada,et al.  Expression of the glucose transporter gene, ptsG, is regulated at the mRNA degradation step in response to glycolytic flux in Escherichia coli , 2001, The EMBO journal.

[32]  J. Plumbridge,et al.  Expression of the phosphotransferase system both mediates and is mediated by Mlc regulation in Escherichia coli , 1999, Molecular microbiology.

[33]  J. Plumbridge,et al.  Expression of ptsG, the gene for the major glucose PTS transporter in Escherichia coli, is repressed by Mlc and induced by growth on glucose , 1998, Molecular microbiology.