Summary
DNA fragments of 3.4 kb containing the gyrB gene were cloned from Escherichia coli KL-16 and from spontaneous nalidixic acid-resistant mutants. The mutations (nal-24 and nal-31) had been determined to be in the gyrB gene by transduction analysis. Nucleotide sequence analysis of the cloned DNA fragments revealed that nal-24 was a G to A transition at the first base of the 426th codon of the gyrB gene, resulting in an amino acid change from aspartic acid to asparagine, and nal-31 was an A to G transition at the first base of the 447th codon, resulting in an amino acid change from lysine to glutamic acid. This indicates that mutations in the gyrB gene are responsible for nalidixic acid resistance.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adachi T, Mizuuchi K, Menzel R, Gellert M (1984) DNA sequence and transcription of the region upstream of the E. coli gyrB gene. Nucleic Acids Res 12: 6389–6395
Bachmann BJ (1972) Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev 36: 525–557
Bachmann BJ (1983) Linkage map of Escherichia coli K-12, Edition 7. Bacteriol Rev 47: 180–230
Blanar MA, Sander SJ, Armengod M-E, Ream LW, Clark AJ (1984) Molecular analysis of the recF gene of Escherichia coli. Proc Natl Acad Sci USA 81: 4622–4626
Cosloy SD, Oishi M (1973) The nature of the transformation process in Escherichia coli K12. Mol Gen Genet 124: 1–10
Cozzarelli NR (1980) DNA gyrase and the supercoiling of DNA. Science 207: 953–960
Gellert M, O'Dea MH, Itoh T, Tomizawa J (1976) Novobiocin and coumermycin inhibit DNA supercoiling catalyzed by DNA gyrase. Proc Natl Acad Sci USA 73: 4474–4478
Gellert M, Mizuuchi K, O'Dea MH, Itoh T, Tomizawa J (1977) Nalidixic acid resistance: a second genetic character involved in DNA gyrase activity. Proc Natl Acad Sci USA 74: 4772–4776
Gellert M (1981) DNA topoisomerases. Annu Rev Biochem 50: 879–910
Hansen FG, Meyenburg KV (1979) Characterization of the dnaA, gyrB and other genes in the dnaA region of the Escherichia coli chromosome on specialized transducing phages λ tna. Mol Gen Genet 175: 135–144
Hansen EB, Hansen FG, Meyenburg KV (1982) The nucleotide sequence of the dnaA gene and the first part of the dnaN gene of Escherichia coli K-12. Nucleic Acids Res 10: 7373–7385
Higgins NP, Peebles CL, Sugino A, Cozzarelli NR (1978) Purification of subunits of Escherichia coli DNA gyrase and reconstitution of enzymatic activity. Proc Natl Acad Sci USA 75: 1773–1777
Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114: 193–197
Inoue S, Ohue T, Yamagishi J, Nakamura S, Shimizu M (1978) Mode of incomplete cross-resistance among pipemidic, piromidic, and nalidixic acids. Antimicrob Agents Chemother 14: 240–245
Inoue S, Yamagishi J, Nakamura S, Furutani Y, Shimizu M (1982) Novel nalidixic acid-resistance mutations relating to DNA gyrase activity. Proceedings of drug resistance in bacteria, 1981, pp 411–414
Kato T, Rothman RH, Clark AJ (1977) Analysis of the role of recombination and repair in mutagenesis of Escherichia coli by UV irradiation. Genetics 87: 1–18
Kimura M, Miki T, Hiraga S, Nagata T, Yura T (1979) Conditionally lethal amber mutations in the dnaA region of the Escherichia coli chromosome that affect chromosome replication. J Bacteriol 140: 825–834
Lampe MF, Bott KF (1985) Genetic and physical organization of the cloned gyrA and gyrB genes of Bacillus subtilis. J Bacteriol 162: 78–84
Lesher GY, Froelich EJ, Gruett MD, Bailey JH, Brundage RP (1962) 1,8-Naphthyridine derivatives. A new class of chemotherapeutic agents. J Med Pharm Chem 5: 1063–1065
Low B (1968) Formation of merodiploids in matings with a class of Rec recipient strains of Escherichia coli K12. Proc Natl Acad Sci USA 60: 160–167
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101: 20–78
Miki T, Kimura M, Hiraga S, Nagata T, Yura T (1979) Cloning and physical mapping of the dnaA region of the Escherichia coli chromosome. J Bacteriol 140: 817–824
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Mizuuchi K, Mizuuchi M, O'Dea MH, Gellert M (1984) Cloning and simplified purification of Escherichia coli DNA gyrase A and B proteins. J Biol Chem 259: 9199–9201
Mizuuchi K, O'Dea MH, Gellert M (1978) DNA gyrase: Subunit structure and ATPase activity of the purified enzyme. Proc Natl Acad Sci USA 75: 5960–5963
Moriya S, Ogasawara N, Yoshikawa H (1985) Structure and function of the replication origin of the Bacillus subtilis chromosome. III. Nucleotide sequence of some 10,000 base pairs in the origin region. Nucleic Acids Res 13: 2251–2265
Murakami A, Inokuchi H, Hirota Y, Ozeki H, Yamagishi H (1980) Characterization of the dnaA gene carried by lambda transducing phage. Mol Gen Genet 180: 235–247
Nakamura S, Minami A, Katae H, Inoue S, Yamagishi J, Takase Y, Shimizu M (1983) In vitro antibacterial properties of AT-2266, a new pyridonecarboxylic acid. Antimicrob Agents Chemother 23: 641–648
Ogasawara N, Moriya S, Yoshikawa H (1985) Structure and function of the region of the replication origin of the Bacillus subtilis chromosome. IV. Transcription of the oriC region and expression of DNA gyrase genes and other open reading frames. Nucleic Acids Res 13: 2267–2279
Ohmori H, Kimura M, Nagata T, Sakakibara Y (1984) Structural analysis of the dnaA and dnaN genes of Escherichia coli. Gene 28: 159–170
Orr E, Staudenbauer WL (1982) Bacillus subtilis DNA gyrase: Purification of subunits and reconstitution of supercoiling activity. J Bacteriol 151: 524–527
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467
Schaus N, O'day K, Peters W, Wright A (1981) Isolation and characterization of amber mutations in gene dnaA of Escherichia coli K-12. J Bacteriol 145: 904–913
Shimizu M, Takase Y, Nakamura S, Katae H, Minami A, Nakata K, Inoue S, Ishiyama M, Kubo Y (1975) Pipemidic acid, a new antibacterial agent active against Pseudomonas aeruginosa: in vivo properties. Antimicrob Agents Chemother 8: 132–138
Smith JT (1984) Awakening the slumbering potential of the 4-quinolone antibacterials. Pharm J 15: 299–305
Sugino A, Peebles CL, Kreuzer KN, Cozzarelli NR (1977) Mechanism of action of nalidixic acid: Purification of Escherichia coli nalA gene product and its relationship to DNA gyrase and a novel nicking-closing enzyme. Proc Natl Acad Sci USA 74: 4767–4771
Wilkie NM, Clements JB, Boll W, Mantei N, Lonsdale D, Weissmann C (1979) Hybrid plasmids containing an active thymidine kinase gene of Herpes simplex virus I. Nucleic Acids Res 7: 859–877
Yamagishi J, Furutani Y, Inoue S, Ohue T, Nakamura S, Shimizu M (1981) New nalidixic acid resistance mutations related to deoxyribonucleic acid gyrase activity. J Bacteriol 148: 450–458
Author information
Authors and Affiliations
Additional information
Communicated by M. Takanami
Rights and permissions
About this article
Cite this article
Yamagishi, Ji., Yoshida, H., Yamayoshi, M. et al. Nalidixic acid-resistant mutations of the gyrB gene of Escherichia coli . Molec Gen Genet 204, 367–373 (1986). https://doi.org/10.1007/BF00331012
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00331012