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Nucleic Acids Research, 1989, Vol. 17, No. 24 10337-10351
© 1989

Articles

Superhelical stress restrained in plasmid DNA during repair synthesis initiated by the UvrA, B and C proteins in vitro

Claude Backendorf*, René Olsthoorn and Pieter van de Putte

Radiation Carcinogenesis Group, Department of Molecular Genetics, Gorlaeus Laboratories, Leiden University PO Box 9502, 2300 RA Leiden, The Netherlands

*To Whom Correspondance Should be addressed

Received October 20, 1989. Revised November 20, 1989. Accepted November 20, 1989.

Purified UvrA, UvrB, UvrC, UvrD, PolA and Lig proteins from Escherichia coli have been used to assess the effect of nucleotide excision repair on the conformation of native negatively supercoiled plasmid DNA in an in vitro test system. The analysis of labeled reaction products on specific gel systems suggests that the Uvr excinuclease has the ability to restrain the superhelical stress in the template DNA during the repair process. This feature, observed in the case of the Uvr system is not found if the repair reaction is initiated by T4 endonuclease V or Micrococcus luteus UV endonuclease.


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