Skip to main content
Log in

The E. coli uvrD gene product is DNA helicase II

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

We have shown that the uvrD gene product, previously identified in maxicell extracts as a 73 kilodalton protein, copurifies with single stranded DNA-dependent ATPase and ATP-dependent DNA helicase activities. This protein is specifically precipitated from maxicell extracts by antibodies raised against DNA helicase II. In order to facilitate purification of the UvrD protein we have sub-cloned the uvrD gene into a plasmid vector in which its transcription is under the control of the phage lambda leftward promoter. Using cells harbouring this recombinant plasmid as a source of elevated levels of the UvrD protein we have purified this protein to homogeneity by a simple, rapid procedure. The purified protein has single stranded DNA-dependent ATPase activity and ATP-dependent DNA halicase activity, and both activities are specifically inactivated by antibodies raised against DNA helicase II. We conclude that DNA helicase II is the uvrD gene product.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Abdel-Monem M, Hoffmann-Berling H (1976) Enzymatic unwinding of DNA. I. Purification and characterization of a DNA-dependent ATPase from Escherichia coli. Eur J Biochem 65:431–440

    Google Scholar 

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

    Google Scholar 

  • Abdel-Monem M, Chanal MC, Hoffmann-Berling H (1977a) DNA unwinding enzyme II of Escherichia coli. I. Purification and characterization of the ATPase activity. Eur J Biochem 79:33–38

    Google Scholar 

  • Abdel-Monem M, Durwald H, Hoffmann-Berling H (1977b) DNA unwinding enzyme II of E. coli. II. Characterization of the DNA unwinding activity. Eur J Biochem 79:39–46

    Google Scholar 

  • Arthur HM, Bramhill D, Eastlake P, Emmerson PT (1982) Cloning of the uvrD gene of E. coli and identification of the product. Gene 19:285–295

    Google Scholar 

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

    Google Scholar 

  • Eichler DC, Lehman IR (1977) On the role of ATP in phosphodiester bond hydrolysis catalysed by the recBC deoxyribonuclease of Escherichia coli. J Biol Chem 252:499–503

    Google Scholar 

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

    Google Scholar 

  • Howard-Flanders P, Bardwell E (1981) Effects of recB21, recF143 and uvrD152 on recombination in lambda bacteriophage-prophage and Hfr by F crosses. J Bacteriol 148:739–743

    Google Scholar 

  • Horiuchi T, Nagata T (1973) Mutations affecting growth of the Escherichia coli cell under a condition of DNA polymerase I deficiency. Mol Gen Genet 123:89–110

    Google Scholar 

  • Kacinsky BM, Rupp WD (1981) A general approach for purifying proteins encoded by cloned genes without using a functional assay: isolation of the uvrA gene product from radiolabelled maxicells. Nucl Acids Res 9:4495–4507

    Google Scholar 

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

    Google Scholar 

  • Lundblad V, Kleckner N (1982) Mutants of E. coli K-12 which affect excision of transposon Tn10. In: Lemont JF, Generoso WM (eds) Molecular and cellular mechanisms of mutagenesis. Plenum, New York

    Google Scholar 

  • Maples VF, Kushner SR (1982) DNA repair in Escherichia coli: Identification of the uvrD gene product. Proc Natl Acad Sci USA 79:5616–5620

    Google Scholar 

  • Messing J, Crea R, Seeburg PH (1981) A system for shotgun DNA sequencing. Nucl Acids Res 9:309–321

    Google Scholar 

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

    Google Scholar 

  • Remaut E, Stanssens P, Fiers W (1981) Plasmid vectors for high efficiency expression controlled by the p l promoter of coliphage lambda. Gene 15:81–93

    Google Scholar 

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

    Google Scholar 

  • Rothman RH (1978) Dimer excision and repair replication patch size in a recL mutant of E. coli K-12. J Bacteriol 136:444–448

    Google Scholar 

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

    Google Scholar 

  • Sancar A, Hack AM, Rupp WD (1979) Simple method for the identification of plasmid-coded proteins. J Bacteriol 137:692–693

    Google Scholar 

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

    Google Scholar 

  • Siegel EC (1973b) Ultraviolet-sensitive mutator mutU4 of Escherichia coli inviable with polA. J Bacteriol 113:161–166

    Google Scholar 

  • Smirnov GB, Filkova EV, Skavronskaya AG (1972) The mutator property of uvr-502 mutation affecting UV-sensitivity of Escherichia coli. Mol Gen Genet 118:51–56

    Google Scholar 

  • Smirnov GB, Filkova EV, Stavronskaya AG (1973) Loss and restoration of viability of E. coli due to combinations of mutations affecting DNA polymerase I and repair activities. Mol Gen Genet 121:139–150

    Google Scholar 

  • Van Sluis CA, Mattern IE, Paterson MC (1974) Properties of uvrE mutants of Escherichia coli K12. I. Effects of UV-irradiation on DNA metabolism Mutat Res 25:273–279

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by D. Sherratt

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hickson, I.D., Arthur, H.M., Bramhill, D. et al. The E. coli uvrD gene product is DNA helicase II. Mol Gen Genet 190, 265–270 (1983). https://doi.org/10.1007/BF00330649

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330649

Keywords

Navigation