DNA polymerase β-dependent long patch base excision repair in living cells

Kenjiro Asagoshi, Yuan Liu, Aya Masaoka, Li Lan, Rajendra Prasad, Julie K. Horton, Ashley R. Brown, Xiao hong Wang, Hussam M. Bdour, Robert W. Sobol, John Stephen Taylor, Akira Yasui, Samuel H. Wilson

    Research output: Contribution to journalArticlepeer-review

    39 Citations (Scopus)

    Abstract

    We examined a role for DNA polymerase β (Pol β) in mammalian long patch base excision repair (LP BER). Although a role for Pol β is well known in single-nucleotide BER, information on this enzyme in the context of LP BER has been limited. To examine the question of Pol β involvement in LP BER, we made use of nucleotide excision repair-deficient human XPA cells expressing UVDE (XPA-UVDE), which introduces a nick directly 5′ to the cyclobutane pyrimidine dimer or 6-4 photoproduct, leaving ends with 3′-OH and 5′-phosphorylated UV lesion. We observed recruitment of GFP-fused Pol β to focal sites of nuclear UV irradiation, consistent with a role of Pol β in repair of UV-induced photoproducts adjacent to a strand break. This was the first evidence of Pol β recruitment in LP BER in vivo. In cell extract, a 5′-blocked oligodeoxynucleotide substrate containing a nicked 5′-cyclobutane pyrimidine dimer was repaired by Pol β-dependent LP BER. We also demonstrated Pol β involvement in LP BER by making use of mouse cells that are double null for XPA and Pol β. These results were extended by experiments with oligodeoxynucleotide substrates and purified human Pol β.

    Original languageEnglish
    Pages (from-to)109-119
    Number of pages11
    JournalDNA Repair
    Volume9
    Issue number2
    DOIs
    Publication statusPublished - 2010 Feb 4

    Keywords

    • DNA polymerase β
    • Flap endonuclease 1
    • Long patch base excision repair
    • Thymine dimer
    • UV damage endonuclease

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

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