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Nucleotide insertion opposite a cis-syn thymine dimer by a replicative DNA polymerase from bacteriophage T7

  • Ying Li
  • , Shuchismita Dutta
  • , Sylvie Doublié
  • , Hussam Moh d. Bdour
  • , John Stephen Taylor
  • , Tom Ellenberger

Research output: Contribution to journalArticlepeer-review

Abstract

Ultraviolet-induced DNA damage poses a lethal block to replication. To understand the structural basis for this, we determined crystal structures of a replicative DNA polymerase from bacteriophage T7 in complex with nucleotide substrates and a DNA template containing a cis-syn cyclobutane pyrimidine dimer (CPD). When the 3′ thymine is the templating base, the CPD is rotated out of the polymerase active site and the fingers subdomain adopts an open orientation. When the 5′ thymine is the templating base, the CPD lies within the polymerase active site where it base-pairs with the incoming nucleotide and the 3′ base of the primer, while the fingers are in a closed conformation. These structures reveal the basis for the strong block of DNA replication that is caused by this photolesion.

Original languageEnglish
Pages (from-to)784-790
Number of pages7
JournalNature Structural and Molecular Biology
Volume11
Issue number8
DOIs
StatePublished - Aug 2004

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