Activity of calf thymus DNA helicase E on cis-diamminedichloroplatinum (II)-damaged DNA

L. Huang, J. J. Turchi, A. F. Wahl, R. A. Bambara

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Calf DNA helicase E (hel E) is a moderately processive, 3' to 5' helicase, active on nicked DNA, that we have proposed to have a role in DNA repair (Turchi, J. J., Murante, R. S., and Bambara, R. A. (1992) Nucleic Acids Res. 20, 6075-6080). Here we have examined its activity on a series of cis- diamminedichloroplatinum (II) (cis-DDP)-modified DNA substrates. Hel E was capable of efficiently displacing a primer strand containing, in an internal position, a cis-DDP-modified dGG. In a two-primer model system, calf DNA polymerase ε could successfully extend an upstream primer through a cis- DDP-modified downstream primer, to the end of the complementary template strand, in a reaction dependent on hel E. However, the translocation of hel E was blocked by cis-DDP modification of the template strand. Primer displacement was completely prevented if the modified site was located just upstream of the primer. The DNA-dependent ATPase activity of helicase E was also reduced by cis-DDP modification of the template DNA. Substrate competition experiments indicated that cis-DDP-modified DNA templates did not sequester hel E. Substrate titration experiments suggested that there is a short delay without ATP hydrolysis before dissociation of helicase E from cis-DDP-modified template sites. Interestingly, hel E could displace a primer if the cis-DDP modification was on the template within the annealed region. Possible explanations for this are discussed. Taken together, these results are consistent with the proposal that hel E participates in DNA repair by displacing segments of damaged DNA.

Original languageEnglish (US)
Pages (from-to)26731-26737
Number of pages7
JournalJournal of Biological Chemistry
Volume268
Issue number35
StatePublished - Jan 1 1993
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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