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Restored DNA-binding of the CAMP-CRP activator complex reestablishes negative regulation by the CytR repressor in the deoP2 promoter in Escherichia coli

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Summary

We have investigated in vivo the coupling between CytR regulation of the deoP2 promoter in Escherichia coli and the DNA-binding specificity of the cAMP-CRP (cAMP receptor protein) complex in order to obtain a more detailed picture of the role played by cAMP-CRP in CytR regulation. By introducing CRP proteins that exhibit an altered DNA binding specificity into a strain containing a mutant deoP2 promoter in which cAMP-CRP activation was decreased and CytR regulation completely abolished, we show that CytR regulation of this promoter can be reestablished by restored the DNA binding of the cAMP-CRP complex. Hence, CytR regulation of deoP2 can be modulated by simply varying DNA binding of cAMP-CRP. These data confirm the crucial role played by the cAMP-CRP activator complex in CytR regulation of the deoP2 promoter.

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References

  • Adhya S (1989) Multipartite genetic control elements: Communication by DNA loop. Annu Rev Genet 23:227–250

    Google Scholar 

  • Aiba H, Fujimoto S, Ozaki N (1982) Molecular cloning and nucleotide sequencing of the gene for E. coli cAMP receptor protein. Nucl Acids Res 10:1345–1361

    Google Scholar 

  • Bolivar F (1978) Construction and characterization of new cloning vehicles. III. Derivatives of plasmid pBR322 carrying unique EcoRI sites for selection of EcoRI-generated recombinant molecules. Gene 4:121–136

    Google Scholar 

  • Bremer E, Gerlach P, Middendorf A (1988) Double negative and positive control of tsx expression in Escherichia coli. J Bacteriol 170:108–116

    Google Scholar 

  • de Crombrugghe B, Busby S, Buc H (1984) Cyclic AMP receptor protein: role in transcription activation. Science 224:831–838

    Google Scholar 

  • Ebright RH, Cossart P, Gicquel-Sanzey B, Beckwith J (1984) Mutations that alter the DNA sequence specificity of the catabolite gene activator protein of E. coli. Nature 311:232–235

    Google Scholar 

  • Ebright RH, Kolb A, Buc H, Kunkel T, Krakow JS, Beckwith J (1987) Role of glutamic acid-181 in DNA sequence recognition by the catabolite gene activator protein (CAP) of Escherichia coli: Altered DNA sequence-recognition properties of vat181CAP and leu181 CAP. Proc Natl Acad Sci USA 84:6083–6087

    Google Scholar 

  • Eichenlaub R, Wehlmann H, Ebbers J (1981) Plasmid mini-F encoded functions involved in replication and incompatibility. In: Levy SB, Clowes RC, Koenig EL (eds) Molecular biology, pathogenicity, and ecology of bacterial plasmids. Plenum Press, New York, pp 327–336

    Google Scholar 

  • Gerlach P, Valentin-Hansen P, Bremer E (1990) Transcriptional regulation of the cytR repressor gene of Escherichia coli: autoregulation and positive control by the cAMP/CRP complex. Mol Microbiol 4:479–488

    Google Scholar 

  • Gralla JD (1989) Bacterial gene regulation from distant DNA sites. Cell 57:193–195

    Google Scholar 

  • Kouzarides T, Ziff E (1988) The role of the leucine zipper in the fos-jun interaction. Nature 336:646–651

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Lovett MA, Helinski DR (1976) Method for the isolation of the replication origin of a bacterial replicon: Construction of a mini-F′km plasmid. J Bacteriol 127:982–987

    Google Scholar 

  • Maniatis T, Fritsch ET, Sambrook G (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Martinussen J, Møllegaard NE, Holst B, Douthwaite SR, ValentinHansen P (1989) A new version of negative control: DNA sequences involved in expression and regulation of CytR and CAMP-CRP controlled genes in Escherichia coli. In: Gralla JD (ed) DNA-protein interactions in transcription. Alan R. Liss, New York, pp 34–41

    Google Scholar 

  • McKay DB, Weber IT, Steitz TA (1982) Structure of catabolite activator protein at 2.9 Å resolution. J Biol Chem 257:9518–9524

    Google Scholar 

  • Miller HJ (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Pabo CO, Sauer RT (1984) Protein-DNA recognition. Annu Rev Biochem 53:293–321

    Google Scholar 

  • Phillips SEV, Manfield I, Parsons I, Davidson BE, Rafferty JB, Somers WS, Margarita D, Cohen GN, Saint-Girons I, Stockley PG (1989) Cooperative tandem binding of met repressor of Escherichia coli. Nature 341:711–715

    Google Scholar 

  • Plumbridge J, Kolb A (1991) CAP and Nag repressor binding to the regulatory regions of the nagE-B and manX genes of Escherichia coli. J Mol Biol 217:661–679

    Google Scholar 

  • Ptashne M (1986) A genetic switch. Blackwell Scientific Publications and Cell Press, Palo Alto, Calif. and Cambridge, Mass

    Google Scholar 

  • Stark MJR (1987) Multicopy expression vectors carrying the lac repressor gene for regulated high-level expression of genes in Escherichia coli. Gene 51:255–267

    Google Scholar 

  • Søgaard-Andersen L, Martinussen J, Møllegaard NE, Douthwaite SR, Valentin-Hansen P (1990a) The CytR repressor antagonizes cAMP-CRP activation in the deoP2 promoter of Escherichia coli-K12. J Bacteriol 172:5706–5713

    Google Scholar 

  • Søgaard-Andersen L, Møllegaard NE, Douthwaite SR, ValentinHansen P (1990b) Tandem DNA-bound cAMP-CRP complexes are required for tanscriptional repression of the deoP2 promoter by the CytR repressor in Escherichia coli. Mol Microbiol 4:1595–1601

    Google Scholar 

  • Søgaard-Andersen L, Pedersen H, Holst B, Valentin-Hansen P (1991) A novel function of the cAMP-CRP complex in Escherichia coli: CAMP-CRP functions as an adaptor for the CytR repressor in the deo operon. Mol Microbiol 5:969–975

    Google Scholar 

  • Taylor JW, Ott J, Eckstein F (1985) The rapid generation of oligonucleotide-directed mutations at high frequency using phosphorothioate-modified DNA. Nucleic Acids Res 13:8765–8785

    Google Scholar 

  • Valentin-Hansen P (1982) Tandem CRP binding sites in the deo operon of E. coli. EMBO J 1:1049–1054

    Google Scholar 

  • Valentin-Hansen P (1985) DNA sequences involved in expression and regulation of DeoR, CytR and cAMP-CRP controlled genes in E. coli. In: Glass RE, Spicek J (eds) Gene manipulation and expression. Croom Helm, London, pp 273–288

    Google Scholar 

  • Valentin-Hansen P, Svenningsen I, Munch-Petersen A, Hammer-Jespersen K (1978) Regulation of the deo operon in E. coli. Mol Gen Genet 159:191–202

    Google Scholar 

  • Valentin-Hansen P, Aiba H, Schümperli D (1982) The structure of tandem regulatory regions in the deo operon of E. coli K12. EMBO J 1:317–322

    Google Scholar 

  • Valentin-Hansen P, Hammer-Jespersen K, Böetius F, Svenningsen I (1984) Structure and function of the intercistronic regulatory deoC-deoA element of E. coli K12. EMBO J 3:179–183

    Google Scholar 

  • Valentin-Hansen P, Love Larsen JE, Hojrup P, Short S, Barbier CS (1986) Nucleotide sequence of the CytR regulatory gene of E. coli K12. Nucleic Acids Res 14:2215–2228

    Google Scholar 

  • Valentin-Hansen P, Holst B, Sogaard-Andersen L, Martinussen J, Nesvera J, Douthwaite SR (1991) Design of cAMP-CRP activated promoters in Escherichia coli. Mol Microbiol 5:433–437

    Google Scholar 

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Communicated by J.W. Lengeler

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Søgaard-Andersen, L., Valentin-Hansen, P. Restored DNA-binding of the CAMP-CRP activator complex reestablishes negative regulation by the CytR repressor in the deoP2 promoter in Escherichia coli . Molec. Gen. Genet. 231, 76–80 (1991). https://doi.org/10.1007/BF00293824

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  • DOI: https://doi.org/10.1007/BF00293824

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