Biochemical Characterization of Uracil-DNA Glycosylase from Pyrococcus furiosus  

作　　者：L1N Li-bo LIU Yu-fen LIU Xi-peng LIU Jian-hua 

机构地区：[1]Key Laboratory of Microbial Metabolism,School of Life Sciences&Biotechnology,Shanghai Jiaotong University,Shanghai 200240,P.R.China

出　　处：《Chemical Research in Chinese Universities》2012年第3期477-482,共6页高等学校化学研究（英文版）

基　　金：Supported by the National High Technology Research and Development Program of China(No.2006AA02Z108);the National Basic Research Program of China(No.2009CB118906);the National Natural Science Foundation of China(Nos.30700131,30870512)

摘　　要：We report the characterization of a uracil-DNA glycosylase（UDG） from the hyperthermophilic archaea Pyrococcus furiosus（P, furiosus）. P. furiosus UDG（PfUDG） has high sequence similarity to the families IV and V UDGs（thermostable UDG family and PaUDG-b family）. PfUDG excises uracil from various DNA substrates with the following order： U/T=U/C〉U/G=U/AP=U/-〉U/U=U/I=U/A. The optimal temperature and pH value for uracil exci- sion by PfUDG are 70 ℃ and 9.0, respectively. The removal of U is inhibited by the divalent ions of Fe, Ca, Zn, Cu, Co, Ni and Mn, as well as a high concentration of NaC1. The phosphorothioates near uracil strongly inhibit the exci- sion of uracil by PfUDG. Interestingly, pfuDNA（Pyrococcusfuriosus DNA） polymerase, which tightly binds the ura- cil-carrying oligonucleotide, does not inhibit the excision by Pfl.IDG, suggesting PfUDG in vivo functions as the re- pair enzyme to excise uracil damage in genome.We report the characterization of a uracil-DNA glycosylase（UDG） from the hyperthermophilic archaea Pyrococcus furiosus（P, furiosus）. P. furiosus UDG（PfUDG） has high sequence similarity to the families IV and V UDGs（thermostable UDG family and PaUDG-b family）. PfUDG excises uracil from various DNA substrates with the following order： U/T=U/C〉U/G=U/AP=U/-〉U/U=U/I=U/A. The optimal temperature and pH value for uracil exci- sion by PfUDG are 70 ℃ and 9.0, respectively. The removal of U is inhibited by the divalent ions of Fe, Ca, Zn, Cu, Co, Ni and Mn, as well as a high concentration of NaC1. The phosphorothioates near uracil strongly inhibit the exci- sion of uracil by PfUDG. Interestingly, pfuDNA（Pyrococcusfuriosus DNA） polymerase, which tightly binds the ura- cil-carrying oligonucleotide, does not inhibit the excision by Pfl.IDG, suggesting PfUDG in vivo functions as the re- pair enzyme to excise uracil damage in genome.

关 键 词：Pyrococcus furiosus（P furiosus） Uracil DNA glycosylase（UDG） Pyrococcus furiosus DNA polymeras Uracil repair in hyperthermophile 

分 类 号：Q959.12[生物学—动物学] Q939.506