base excision by thymine dna glycosylase mediates dna-directed cytotoxicity of 5-fluorouracil基本切除胸腺嘧啶dna糖基化酶介导dna-directed 5 -氟尿嘧啶的细胞毒性.pdfVIP

PLoS BIOLOGY Base Excision by Thymine DNA Glycosylase Mediates DNA-Directed Cytotoxicity of 5-Fluorouracil 1 1 1 1 2 3 ¨ 1* Christophe Kunz , Frauke Focke , Yusuke Saito , David Schuermann , Teresa Lettieri , Jim Selfridge , Primo Schar ¨ ¨ 1 Institute of Biochemistry and Genetics, Department of Biomedicine, University of Basel, Basel, Switzerland, 2 Institute of Cell Biology, ETH Zurich, Zurich, Switzerland, 3 Wellcome Trust Centre for Cell Biology, Institute of Cell and Molecular Biology, University of Edinburgh, Edinburgh, Scotland 5-Fluorouracil (5-FU), a chemotherapeutic drug commonly used in cancer treatment, imbalances nucleotide pools, thereby favoring misincorporation of uracil and 5-FU into genomic DNA. The processing of these bases by DNA repair activities was proposed to cause DNA-directed cytotoxicity, but the underlying mechanisms have not been resolved. In this study, we investigated a possible role of thymine DNA glycosylase (TDG), one of four mammalian uracil DNA glycosylases (UDGs), in the cellular response to 5-FU. Using genetic and biochemical tools, we found that inactivation of TDG significantly increases resistance of both mouse and human cancer cells towards 5-FU. We show that excision of DNA-incorporated 5-FU by TDG generates persistent DNA strand breaks, delays S-phase progression, and activates DNA damage signaling, and that the repair of 5-FU–induced DNA strand breaks is more efficient in the absence of TDG. Hence, excision of 5-FU by TDG, but not by othe