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Escherichia coli uvrD mutants with thermosensitive DNA-dependent Adenosine triphosphatase I (Helicase II)

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Summary

Three mutants producing thermosensitive DNA-dependent Adenosine triphosphatase (ATPase) I were screened from a collection of temperature-sensitive mutants of Escherichia coli K12. ATPase I purified to near homogeneity from one of the mutants (JE11000) possesses both thermosensitive DNA-dependent ATPase and DNA helicase activities. We have shown that ATPase I is encoded by the uvrD gene as first suggested by Oeda et al. (1982): (i) the thermosensitive ATPase I mutation present in JE11040 lies in or very close to the uvrD gene, (ii) ATPase I activity is absent in uvrD210, uvrD156, and uvrD252 mutants. Thus the thermosensitive mutations correspond to new uvrD mutations. However, the mutation present in JE11040 confers neither UV sensitivity nor mutator phenotype at high temperature. Evidence is presented that the mutant ATPase I is stabilized in vivo at 42°C.

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References

  • Abdel-Monem M, Dürwald H, Hoffmann-Berling H (1976) Enzymatic unwinding of DNA. 2. Chain separation by an ATP-dependent DNA unwinding enzyme. Eur J Biochem 65:441–449

    Google Scholar 

  • Abdel-Monem M, Dürwald H, Hoffmann-Berling (1977) DNA unwinding enzyme II of Escherichia coli. 2. Characterization of the DNA unwinding activity. Eur J Biochem 79:39–45

    Google Scholar 

  • Arthur HM, Lloyd RG (1980) Hyper-recombination in uvrD mutants of Escherichia coli K-12. Molec Gen Genet 180:185–191

    Google Scholar 

  • Geider K, Hoffmann-Berling H (1981) Proteins controlling the helical structure of DNA. Ann Rev Biochem 50:233–260

    Google Scholar 

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

    Google Scholar 

  • Hanawalt PC, Cooper PK, Ganesan AK, and Smith CA (1979) DNA repair in bacteria and mammalian cells. Ann Rev Biochem 48:783–836

    Google Scholar 

  • Hickson ID, Arthur HM, Bramhill D, and Emmerson PT (1983) The E. coli uvrD gene product is DNA helicase II. Mol Gen Genet 190:265–270

    Google Scholar 

  • Horii ZI, Clark AJ (1973) Genetic analysis of the recF pathway to genetic recombination in Escherichia coli K-12: isolation and characterization of mutants. J Mol Biol 80:327–344

    Google Scholar 

  • Isono K, Krauss J, and Hirota Y (1976) Isolation and characterization of temperature-sensitive mutants of Escherichia coli with altered ribosomal proteins. Mol Gen Genet 149:297–302

    Google Scholar 

  • Kleckner N (1981) Transposable elements in prokaryotes. Ann Rev Genet 15:341–404

    Google Scholar 

  • Klinkert MQ, Klein A, and Abdel-Monem M (1980) Studies on the functions of DNA helicase I and DNA helicase II of Escherichia coli. J Biol Chem 255:9746–9752

    Google Scholar 

  • Kuemmerle NB, Masker WE (1980) Effect of the uvrD mutation on excision repair. J Bacteriol 142:535–546

    Google Scholar 

  • Kuhn B, Abdel-Monem M (1982) DNA synthesis at a fork in the presence of DNA helicases. Eur J Biochem 125:63–68

    Google Scholar 

  • Kuhn B, Abdel-Monem M, Krell H, and Hoffmann-Berling H (1979) Evidence for two mechanisms for DNA unwinding catalysed by DNA helicases. J Biol Chem 254:11343–11350

    Google Scholar 

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

    Google Scholar 

  • Lane DHE, Denhardt DT (1975) The rep mutation IV. Slower movement of replication forks in Escherichia coli rep strains. J Mol Biol 97:99–112

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, and Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

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

    Google Scholar 

  • Nevers P, Spatz HC (1975) Escherichia coli mutants uvrD and uvrE deficient in gene conversion of λ-heteroduplexes. Mol Gen Genet 139:233–248

    Google Scholar 

  • Oeda K, Horiuchi T, and Sekiguchi M (1981) Molecular cloning of the uvrD gene of Escherichia coli that controls ultraviolet sensitivity and spontaneous mutation frequency. Mol Gen Genet 184:191–199

    Google Scholar 

  • Oeda K, Horiuchi T, and Sekiguchi M (1982) The uvrD gene of E. coli encodes a DNA-dependent ATPase. Nature 298:98–100

    Google Scholar 

  • Radman M, Wagner RE, Glickman BW, and Meselson M (1980) DNA methylation, mismatch correction and genetic stability; In: Alcevic M (ed) Progress in environmental mutagenesis, Elsevier, pp 121–130

  • Reichardt L (1972) PhD Thesis, Standford University

  • Richet E, Kohiyama M (1976) Purification and characterization of a DNA-dependent ATPase from Escherichia coli. J Biol Chem 251:808–812

    Google Scholar 

  • Richet E, Moreau J, and Kohiyama M (1979) DNA-dependent ATPases from Escherichia coli K12. Cold Spring Harbor Symp Quant Biol 43:345–348

    Google Scholar 

  • Richet E, Kern R, Kohiyama M, and Hirota Y (1980) Isolation of DNA-dependent ATPase I mutants of E. coli. In: Alberts B (ed) Mechanistic studies of DNA replication and genetic recombination, vol 19, Academic press, New-York, pp 605–608

    Google Scholar 

  • Rothman RH, Clark AJ (1977) Defective excision and postreplication repair of UV-damaged DNA in a recL mutant strain of E. coli K-12. Mol Gen Genet 155:267–277

    Google Scholar 

  • Sancar A, Rupp WD (1983) A novel repair enzyme: UVRABC excision nuclease of Escherichia coli cuts a DNA strand on both sides of the damaged region. Cell 33:249–260

    Google Scholar 

  • Schaffner W, Weissmann C (1973) A rapid, sensitive and specific method for the determination of protein in dilute solution. Anal Biochem 56:502–514

    Google Scholar 

  • Scott JF, Eisenberg S, Bertsch LI, and Kornberg A (1977) A mechanism of duplex DNA replication revealed by enzymatic studies of phage ΦX174: Catalytic strand separation in advance of replication. Proc Natl Acad Sci USA 74:193–197

    Google Scholar 

  • Scott JF, Kornberg A (1978) Purification of the rep protein of Escherichia coli. J Biol Chem 253:3292–3297

    Google Scholar 

  • Siegel EC (1973) Ultraviolet-sensitive mutator strain of Escherichia coli K-12. J Bacteriol 113:145–160

    Google Scholar 

  • Siegel EC (1981) Complementation studies with the repair-deficient uvrD3, uvrE156 and recL152 mutations in Escherichia coli. Mol Gen Genet 184:526–530

    Google Scholar 

  • Yarranton GT, Das RH, Gefter ML (1979) Enzyme-catalysed DNA unwinding. Mechanism of action of helicase III. J Biol Chem 254:12002–12006

    Google Scholar 

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Communicated by T. Yura

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Richet, E., Nishimura, Y., Hirota, Y. et al. Escherichia coli uvrD mutants with thermosensitive DNA-dependent Adenosine triphosphatase I (Helicase II). Mol Gen Genet 192, 378–385 (1983). https://doi.org/10.1007/BF00392178

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