Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Isolation and characterization of Escherichia coli antimutators

A new strategy to study the nature and origin of spontaneous mutations

  • 56 Accesses

  • 29 Citations


To identify the nature and origin of spontaneous mutability we developed a screening procedure suitable to isolate antimutators showing a lower error rate than 10-10 per base per replication. Among about 500,000 mutagenized colonies we found 20 mutants showing a reduced spontaneous mutability. These antimutators can be subdivided into three groups: (i) Mutants in which the level of spontaneous mutability.is reduced due to an increase in efficiency of the error correcting mechanism (amu4). (ii) Mutants which are deficient in several pathways of DNA repair. This finding supports the hypothesis that much spontaneous mutability is due to error-prone repair (amu59, amu47, amu50, amu62, amu43, amu38). (iii) Mutants in which the antimutator effect seems to be the result of an auxotrophy such as Pur- (amu17), Thr- (amu1, amu28) and Ser- (amu31). This finding might support the hypothesis that metabolically induced lesions are important in spontaneous mutagenesis.

Eleven of these antimutators were mapped at ten bacterial loci in the following positions: amu31 (2 min); amu4 (4 min); amu62 (82 min); amu47 (85 min); amu59 (86 min); amu17 (89 min); amu50 (95 min); amu1/amu28 (100 min); amu38 (23–27 min) and amu43 (74–81 min).

This is a preview of subscription content, log in to check access.


  1. Bachmann B (1983) Linkage map of Escherichia coli K-12, edn 7. Microbiol Rev 47:180–230

  2. Bessman M, Muzyczka N, Goodman N, Schnaar RL (1974) Studies on the biochemical basis of spontaneous mutation. II. J Mol Biol 88:409–421

  3. Cilento G (1980) Phytochemistry in the dark. Photochem Photobiol Rev 5:199–228

  4. Clarke GH, Shankel DM (1975) Antimutagenesis in microbiol systems. Bacteriol Rev 39:33–53

  5. Cox EC (1976) Bacterial mutator genes and the control of spontaneous mutation. Ann Rev Genet 10:135–156

  6. Cox EC, Horner DL (1983) Structure and coding properties of a dominant Escherichia coli mutator gene, mutD. Proc Natl Acad Sci USA 80:2295–2299

  7. Craig N, Roberts J (1981) Function of nucleoside triphosphate and polynucleotide in E. coli recA protein-directed cleavage of lambda repressor. J Biol Chem 256:8039–8044

  8. Drake JW (1969) Spontaneous mutation. Comparative rates of spontaneous mutation. Nature 221:1128–1132

  9. Echols H (1982) Mutationsrate: some biological and biochemical considerations. Biochimie 64:571–575

  10. Echols H, Lu C, Burgers PM (1983) Mutators strains of E. coli, mutD and dnaQ, with defective exonucleolytic editing by DNA polymerase III holoenzyme. Proc Natl Acad Sci USA 80:2189–2192

  11. Freese E (1959) The specific mutagenic effect of base analogues on phage T4. J Mol Biol J 1:87–105

  12. Geiger JR, Speyer JF (1977) A conditional antimutator of E. coli. Mol Gen Genet 153:87–97

  13. Glickman B, Radman M (1980) E. coli mutator mutants deficient in methylation-instructed mismatch correction. Proc Natl Acad Sci USA 77:1063–1067

  14. Glickman B, Van den Elsen P, Radman M (1978) Induced mutagenesis in dam- mutants of E. coli: A role for 6-methyladenine residues in mutation avoidance. Mol Gen Genet 163:307–312

  15. Hall RM, Brammer WJ (1973) Increased spontaneous mutation rates in mutants with altered DNA polymerase III. Mol Gen Genet 121:271–276

  16. Hartmann PE (1980) Bacterial mutagenesis: review of new insights. Environmental Mutagenesis 2:3–16

  17. Hopkins DR, Goodman MF (1980) Desoxyribonucleotide pools, base pairing and sequence configuration affecting bromdeoxyuridine and 2-Aminopurine-induced mutagenesis. Proc Natl Acad Sci USA 77:1801–1805

  18. Horiuchi T, Maki H, Sekuguchi M (1978) A new conditional lethal mutator (dnaQ49) in E. coli. Mol Gen Genet 163:277–283

  19. Kelley W, Chalmers K, Murray N (1977) Isolation and characterization of a Lambda-polA transducing phage. Proc Natl Acad Sci USA 74:5632–5636

  20. Kondo S, Ichikawa H, Iwo K, Kato T (1970) Basechange mutagenesis and prophage induction in strains of E. coli with different repair capacities. Genetics 66:187–217

  21. Konrad E (1978) Isolation of an E. coli K-12 dnaE Mutation as a mutator. J Bacteriol 133:1197–1202

  22. Lederberg J (1952) Allelic relationship and reverse mutation in E. coli. Genetics 37:469–483

  23. Little JW, Mount DW (1982) The SOS regulatory system of E. coli. Cell 29:11–22

  24. Low KB (1973) Rapid mapping of conditional and auxotrophic mutations in E. coli. J Bacteriol 113:798–812

  25. Maenhaut-Michel M, Caillet-Fauquet P (1982a) 2-Aminopurine induced DNA repair in E. coli. Mol Gen Genet 188:143–148

  26. Maenhaut-Michel M, Caillet-Fauquet P (1982b) Effects of E. coli mutator genes mutH, mutL and mutS on 2-Aminopurine induced DNA repair. Biochimie 64:739–742

  27. Marinus M, Morris N (1973) Isolation of DNA methylase mutants of E. coli K-12. J Bacteriol 114:1143–1150

  28. Mergeay M, Faelen M, Gerits I, Lejenne P (1981) Mapping of antimutator mutations in Salmonella typhimurium. Arch Int Physiol Biochem 89:71–72

  29. Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York

  30. Nevers P, Spatz H (1975) E. coli mutants uvrD and uvrE deficient in gene conversion of lambda heteroduplexes. Mol Gen Genet 139:233–243

  31. Persing DH, McGinty L, Adams C, Fowler RG (1981) Mutational specificity of the base analoge 2-aminopurine. Mutat Res 83:25–37

  32. Phizicky EM, Roberts JW (1981) Induction of SOS functions: regulation of proteolytic activity of E. coli RecA protein by interaction with DNA and nucleoside triphosphate. Cell 25:259–267

  33. Quiñones A (1984) Isolation und Charakterisierung von Antimutatormutanten von E. coli. Dissertation, Martin-Luther-University Halle, GDR

  34. Quiñones A, Piechocki R (1981) A direct selection method for isolating antimutators of E. coli. Z Allg Microbiol 21:57–61

  35. Ronen A (1979) 2-Aminopurine. Mutat Res 75:1–47

  36. Ryan FJ, Schwartz M, Fried P (1955) The direct enumeration of spontaneous and induced mutations in bacteria. J Bacteriol 69:552–556

  37. Rydberg B (1978) Bromuracil mutagenesis and mismatch repair in mutator strains of E. coli. Mutat Res 52:11–24

  38. Sargentini NJ, Smith KC (1981) Much of spontaneous mutagenesis in E. coli is due to error-prone DNA repair: implications for spontaneous carcinogenesis. Carcinogenesis 2:863–872

  39. Scheuerman R, Tam S, Burgers PM, Echols H (1983) Identification of the ɛ-subunit of E. coli DNA polymerase III holoenzyme as the dnaQ gene product: a fidelity subunity for DNA replication. Proc Natl Acad Sci USA 80:7085–7089

  40. Schnaar RL, Muzyczka N, Bessman M (1973) Utilisation of 2-aminopurine deoxynucleoside triphosphate by mutator, antimutator and wild-type DNA polymerase of T4. Genetics, Suppl 73:137–140

  41. Sevastopoulos C, Glaser DA (1977) Mutator action by E. coli strains carrying dnaE mutations. Proc Natl Acad Sci USA 74:3947–3950

  42. Sevastopoulos C, Wehr C, Glaser DA (1977) Large-scale automated isolation of E. coli mutants with thermosensitive DNA replication. Proc Natl Acad Sci USA 74:3485–3489

  43. Stacey K, Oliver P (1977) A novel pleiotropic mutation in E. coli which affects transduction, transformation and rates of mutation. J Gen Microbiol 98:569–578

  44. Volkert MR, Nguyen DC (1984) Induction of specific E. coli genes by sublethal treatments with alkylating agents. Proc Natl Acad Sci USA 81:4110–4114

  45. Witkin EM (1976) Ultraviolet mutagenesis and inducible DNA repair in E. coli. Bacteriol Rev 40:869–907

  46. Zamenhof P (1969) On the indentity of two bacterial mutator genes: effect of antimutagens. Mutat Res 7:463–465

Download references

Author information

Correspondence to Ariel Quiñones.

Additional information

Communicated by H. Böhme

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Quiñones, A., Piechocki, R. Isolation and characterization of Escherichia coli antimutators. Molec. Gen. Genet. 201, 315–322 (1985). https://doi.org/10.1007/BF00425677

Download citation


  • Escherichia Coli
  • Error Rate
  • Lower Error
  • Spontaneous Mutability
  • Screen Procedure