Summary
We have studied the correlation between the activities of adenylate cyclase (ATP pyrophosphatelyase-(cyclizing); EC 4.6.1.1) and in vivo rates of synthesis and intracellular concentrations of adenosine 3′,5′ cyclic monophosphate (cAMP) under various growth conditions in wild-type Escherichia coli and in mutants lacking or overproducing the cAMP receptor protein (CAP). We showed that when wild-type bacteria are grown in the presence of a variety of carbon sources the intracellular concentrations of cAMP are inversely related to the adenylate cyclase activities determined in permeabilized cells, suggesting that the carbon source-dependent modulation of cAMP levels is not directly related to the regulation of adenylate cyclase activity. In mutants lacking functional CAP (crp) the in vivo rates of cAMP synthesis are several hundred-fold higher than in the wild-type parent without a parallel increase of adenylate cyclase activities. In a strain carrying multiple copies of the crp gene and overproducing CAP the activity of adenylate cyclase is severely inhibited, although the in vivo rate of cAMP synthesis is similar to the parental strain. We interpret these results as indicating that CAP controls mainly the activity rather than the synthesis of adenylate cyclase.
Similar content being viewed by others
References
Botsford JL, Drexler M (1978) The cyclic 3′,5′ adenosine monophosphate receptor protein and regulation of cyclic 3′,5′, adenosine monophosphate synthesis in Escherichia coli. Mol Gen Genet 165:47–56
Cossart P, Gicquel-Sanzey B (1982) Cloning and sequence of the crp gene of Escherichia coli K12. Nucl Acids Res in press
Danchin A, Dondon L, Joseph E, Ullmann A (1981) Transcriptiontranslation coupling and polarity: A possible role of cyclic AMP. Biochimie 63:419–424
Dicou EL, Brachet Ph (1979) Multiple forms of an extracellular cyclic AMP phosphodiesterase from Dictypstelium discoideum. Biochim Biophys Acta 578:232–242
Epstein W, Rothman-Denes LB, Hesse J (1975) Adenosine 3′,5′ cyclic monophosphate as mediator of catabolite repression in Escherichia coli. Proc Natl Acad Sci USA 72:2300–2304
Feucht BU, Saier MH Jr (1980) Fine control of adenylate cyclase by the phosphoenolpyruvate: sugar phosphotransferase systems in Escherichia coli and Salmonella thyphimurium. J Bacteriol 141:603–610
Frandsen EK, Krishna L (1976) A simple ultrasensitive method for the assay of cyclic AMP and cyclic GMP in tissues. Life Sci 18:529–542
Fraser ADE, Yamazaki H (1978) Determination of the rates of synthesis and degradation of adenosine 3′,5′ cyclic monophosphate in Escherichia coli K 12 CRP- and CRP+ strains. Can J Biochem 56:849–852
Fraser ADE, Yamazaki H (1979) Effect of carbon source on the rates of cyclic AMP synthesis, excretion, and degradation, and the ability to produce β-galactosidase in Escherichia coli. Can J Biochem 57:1073–1079
Guidi-Rontani C, Danchin A, Ullmann A (1980) Catabolite repression in Escherichia coli mutants lacking cyclic AMP receptor protein. Proc Natl Acad Sci USA 77:5799–5801
Guidi-Rontani C, Danchin A, Ullmann A (1981) Isolation and characterization of a mutant affected in the regulation of adenylate cyclase of Escherichia coli. J Bacteriol 148:753–761
Guiso N, Blazy B (1980) Regulatory aspects of the cyclic AMP receptor protein in Escherichia coli K12. Biochem Biophys Res Commun 94:278–283
Harwood JP, Peterkofsky A (1975) Glucose-sensitive adenylate cyclase in toluene-treated cells of Escherichia coli B. J Biol Chem 250:4656–4662
Janeček J, Náprstek J, Dobrová Z, Jirešová M, Spížek J (1979) Adenylate cyclase activity in Escherichia coli cultured under various conditions. FEMS Microbiol Lett 6:305–307
Leive L (1968) Studies of the permeability change produced in coliform bacteria by ethylenediaminetetraacetate. J Biol Chem 243: 32373–2380
Majerfeld IH, Miller D, Spitz E, Rickenberg HV (1981) Regulation of the synthesis of adenylate cyclase in Escherichia coli by the cAMP-cAMP receptor protein complex. Mol Gen Genet 181: 470–475
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor, New York, pp 201–205
Pardee AB, Jacob F, Monod J (1959) The genetic control and cytoplasmic expression of inducibility in the synthesis of β-galactosidase of Escherichia coli. J Mol Biol 1:165–168
Peterkofsky A (1976) Cyclic Nucleotides in Bacteria. Adv Cyclic Nucleotide Res 7:3–47
Peterkofsky A, Gazdar C (1975) Interaction of enzyme I of the phosphoenol-pyruvate sugar: phosphotransferase system with adenylate cyclase. Proc Natl Acad Sci USA 72:2920–2924
Peterkofsky A, Gazdar C (1978) The Escherichia coli adenylate cyclase complex: activation by phosphoenolpyruvate. J Supramol Struct 9:219–230
Postma PW, Schuitema A, Kwa C (1981) Regulation of methyl-β-galactoside permease activity in pts and crr mutants of Salmonella typhimurium. Mol Gen Genet 181:448–453
Potter K, Chaloner-Larsson G, Yamazaki H (1974) Abnormally high rate of cyclic AMP excretion from an Escherichia coli mutant deficient in cyclic AMP receptor protein. Biochem Biophys Res Commun 48:169–174
Rephaeli AW, Saier MH Jr (1976) Effects of crp mutations on adenosine 3′,5′ monophosphate metabolism in Salmonella typhimurium. J Bacteriol 127:120–127
Sabourin D, Beckwith J (1975) Deletion of the Escherichia coli crp genes. J Bacteriol 122:338–340
Saier MH Jr (1977) Bacterial phosphoenolpyruvate: sugar phosphotransferase systems: structural, functional and evolutionary interrelationships. Bacteriol Rev 41:856–871
Saier MH Jr, Feucht BU, McCaman MT (1975) Regulation of intracellular adenosine cyclic 3′,5′ monophosphate levels in Escherichia coli and Salmonella typhimurium. J Biol Chem 250:7593–7601
Scholte BJ, Postma PW (1980) Mutation in the crp gene of Salmonella thyphimurium hwich interferes with inducer exclusion. J Bacteriol 141:751–757
Steiner AL, Wehman RE, Parker CW, Kipnis DM (1972) Radioimmunoassay for the measurement of cyclic nucleotides. Adv Cyclic Nucleotide Res 2:51–61
Ullmann A, Danchin A (1982) Role of cyclic AMP in bacteria. Adv Cyclic Nucleotide Res 15: in press
Wang JYJ, Clegg DO, Koshland E Jr (1981) Molecular cloning and amplification of the adenylate cyclase gene. Proc Natl Acad Sci USA 78:4684–4688
Wayne PK, Rosen OM (1971) Cyclic 3′,5′ adenosine monophosphate in Escherichia coli during transient and catabolite repression. Proc Natl Acad Sci USA 71:1436–1440
Author information
Authors and Affiliations
Additional information
Communicated by G. O'Donovan
Rights and permissions
About this article
Cite this article
Joseph, E., Bernsley, C., Guiso, N. et al. Multiple regulation of the activity of adenylate cyclase in Escherichia coli . Molec. Gen. Genet. 185, 262–268 (1982). https://doi.org/10.1007/BF00330796
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00330796