Summary
We isolated strains of Escherichia coli K 12 in which the lac structural genes were fused to the structural genes of the guaBA operon. These strains were used to isolate regulatory mutations that increased the expression of the guaBA operon under normal repressing conditions as compared to the wild type parental fusion strain. Three classes of guaBA specific regulatory mutations were identified. Class I regulatory mutations were trans-acting and unlinked to the guaBA operon as shown by bacteriophage P1 transduction. Class II regulatory mutations were tightly linked to the guaBA operon, cis-dominant to the wild type allele in a cis-trans analysis and were regarded as control region mutations. Class III regulatory mutations were tightly linked to the guaBA operon and trans-recessive to the wild type allele in a cis-trans analysis. We have designated the locus responsible for the class III regulatory mutations as guaR. The guaR locus is tightly linked and was mapped to the counterclockwise side of the guaBA operon. The guaR locus is proposed to specify a trans acting regulatory element involved in the regulation of the guaBA operon.
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References
Bachmann BJ (1983) Linkage map of Escherichia coli K-12. Ed 7. Microbiol Rev 47:180–230
Benson CE, Gots JS (1976) Occurrence of a regulatory deficiency in purine biosynthesis among purA mutants of Salmonella typhimurium. Mol Gen Genet 148:31–36
Bertani G (1951) Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol 62:293–300
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye-binding. Anal Biochem 72:248–254
Casadaban MJ, Cohen S (1979) Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci USA 76:4530–4533
Datta N, Hedges RW, Shaw EJ, Sykes RB, Richmond MH (1971) Properties of an R factor from Pseudomonas aeruginosa. J Bacteriol 108:1244–1249
Eisenbeis SJ, Parker J (1981) Strains of Escherichia coli carrying the structural gene for histidyl-tRNA synthetase on a highcopy-number plasmid. Mol Gen Genet 183:115–122
Fukumaki Y, Shimada K, Takagi Y (1977) Secondary promoter of the guanine operon of Escherichia coli K12. J Bacteriol 131:685–688
Faelen M, Toussaint A, Van Montagu M, Van den Elsacker S, Engler G, Schell J (1976) In vivo genetic engineering: The Mumediated transposition of chromosomal DNA segments onto transmissible plasmids. In: Bukhari AL, Shapiro JA, Adhya SL (eds) DNA insertion elements, plasmids, and episomes. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 521–530
Gallant J, Irr J, Cashel M (1971) The mechanism of amino acid control of guanylate and adenylate biosynthesis. J Biol Chem 246:5812–5816
Garber BB, Jochimsen BU, Gots JS (1980) Glutamine and related analogs regulate guanosine monophosphate reductase in Salmonella typhimurium. J Bacteriol 143:105–111
Gilbert HJ, Lowe CR, Drabble WT (1979) Inosine 5′-monophosphate dehydrogenase of Escherichia coli: purification by affinity chromatography, subunit structure and inhibition by guanosine 5′-monophosphate. Biochem J 183:481–494
Gots JS, Benson CE, Jochimsen B, Koduri KR (1977) Microbial models and regulatory elements in the control of purine metabolism. In: Purine and Pyrimidine Metabolism. CIBA foundation Symposium. vol 48. Elsevier Press, Amsterdam, pp 23–41
Gowrishankar J, Pittard J (1982) Construction from Mud1 (lacApr) lysogens of lambda bacteriophage bearing promoter-lac fusions: isolation of ppheA-lac. J Bacteriol 150:1122–1129
Houlberg U, Jensen KF (1983) Role of hypoxanthine and guanine in regulation of Salmonella typhimurium pur gene expression. J Bacteriol 153:837–845
Howe MM, Bade EG (1975) Molecular biology of bacteriophage Mu. Science 190:624–632
Koduri RK, Gots JS (1980) A DNA-binding protein with specificity for pur genes in Escherichia coli. J Biol Chem 255:9594–9598
Kuramitsu HK, Udaka S, Moyed HS (1964) Induction of inosine 5′-phosphate dehydrogenase and xanthosine 5′-phosphate aminase by ribosyl-4-amino-5-imidazolecarboxamide in purine requiring mutants of Escherichia coli B. J Biol Chem 239:3425–3430
Lambden PR, Drabble WT (1973) The gua operon of Escherichia coli K-12: evidence for polarity from guaB to guaA. J Bacteriol 115:992–1002
Levin AP (1967) Alteration of repression in purine biosynthesis. Biochem Biophys Acta 138:221–229
Ljungquist E, Bukhari AI (1979) Behavior of bacteriophage Mu DNA upon infection of Escherichia coli cells. J Mol Biol 133:339–357
Mager J, Magasanik B (1960) Guanosine 5′-phosphate reductase and its role in the interconversion of purine nucleotides. J Biol Chem 235:1474–1478
Mehra RK, Drabble WT (1981) Dual control of the gua operon of Escherichia coli K 12 by adenine and guanine nucleotides. J Gen Microbiol 123:27–37
Michaels G, Kelln RA (1983) Construction and use of pyr::lac fusion strains to study regulation of pyrimidine biosynthesis in Salmonella typhimurium. Mol Gen Genet 189:463–470
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Miozzari GF, Niederberger P, Hutter R (1978) Permeabilization of microorganisms by Triton X-100. Anal Biochem 90:220–233
Neidhardt FC, Bloch PL, Smith DF (1974) Culture medium for enterobacteria. J Bacteriol 119:736–747
Nierlich DP, Magasanik B (1971) Control by feedback repression of the enzymes of purine biosynthesis in Aerobacter Aerogenes. Biochem Biophys Acta 230:349–361
Nijkamp HJJ (1969) Regulatory role of adenine nucleotides in the biosynthesis of guanosine 5′-monophosphate. J Bacteriol 100:585–593
Nijkamp HJJ, De Haan PG (1967) Genetic and biochemical studies of the guanosine 5′-monophosphate pathway in Escherichia coli. Biochem Biophys Acta 145:31–40
Rosner JL (1972) Formation, induction and curing of bacteriophage P1 lysogens. Virology 48:679–689
Smith JM, Gots JS (1980) PurF-lac fusion and direction of purF transcription in Escherichia coli. J Bacteriol 143:1156–1164
Smith JM, Smith FJ, Umbarger HE (1979) Mutations affecting the formation of acetohydroxy acid synthase II in Escherichia coli strain K-12. Mol Gen Genet 148:111–124
Spector T (1978) Refinment of Coomassie Blue method of protein quantitation-sample and linear spectrophotometric assay for less-than-or-equal-to 0.5 to 50 microgram of protein. Anal Biochem 86:142–146
Spibey N, Drabble WT (1981) Construction and characterization of guaB-lacZ fusions in Escherichia coli K12. J Gen Microbiol 126:497–501
Thomulka KW, Gots JS (1982) Isolation and characterization of purine regulatory mutants of Salmonella typhimurium with an episomal purE-lac fusion. J Bacteriol 151:153–161
Udaka S, Moyed HS (1963) Inhibition of parental and mutant xanthosine 5′-phosphate aminases by psicofuranine. J Biol Chem 238:2979–2803
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Communicated by A. Bukhari
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Tiedeman, A.A., Smith, J.M. Isolation and characterization of regulatory mutations affecting the expression of the guaBA operon of Escherichia coli K-12. Mol Gen Genet 195, 77–82 (1984). https://doi.org/10.1007/BF00332727
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DOI: https://doi.org/10.1007/BF00332727