Structural insight into dynamic bypass of the major cisplatin-DNA adduct by Y-family polymerase Dpo4

Jimson H.Y. Wong, Jessica A. Brown, Zucai Suo, Paul Blum, Takehiko Nohmi, Hong Ling

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Y-family DNA polymerases bypass Pt-GG, the cisplatin-DNA double-base lesion, contributing to the cisplatin resistance in tumour cells. To reveal the mechanism, we determined three structures of the Y-family DNA polymerase, Dpo4, in complex with Pt-GG DNA. The crystallographic snapshots show three stages of lesion bypass: the nucleotide insertions opposite the 3′G (first insertion) and 5′G (second insertion) of Pt-GG, and the primer extension beyond the lesion site. We observed a dynamic process, in which the lesion was converted from an open and angular conformation at the first insertion to a depressed and nearly parallel conformation at the subsequent reaction stages to fit into the active site of Dpo4. The DNA translocation-coupled conformational change may account for additional inhibition on the second insertion reaction. The structures illustrate that Pt-GG disturbs the replicating base pair in the active site, which reduces the catalytic efficiency and fidelity. The in vivo relevance of Dpo4-mediated Pt-GG bypass was addressed by a dpo-4 knockout strain of Sulfolobus solfataricus, which exhibits enhanced sensitivity to cisplatin and proteomic alterations consistent with genomic stress.

Original languageEnglish (US)
Pages (from-to)2059-2069
Number of pages11
JournalEMBO Journal
Volume29
Issue number12
DOIs
StatePublished - Jun 16 2010

Keywords

  • Anti-cancer drug
  • Cisplatin
  • DNA damage
  • Translesion DNA synthesis
  • Y-family DNA polymerase

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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