Biochemical Characterization of Uracil-DNA Glycosylase from Pyrococcus furiosus

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

作者机构：Key Laboratory of Microbial MetabolismSchool of Life Sciences&BiotechnologyShanghai Jiaotong UniversityShanghai 200240P.R.China 

出 版 物：《Chemical Research in Chinese Universities》 (高等学校化学研究（英文版）)

年 卷 期：2012年第28卷第3期

页      面：477-482页

核心收录：

学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 07[理学] 071005[理学-微生物学] 071002[理学-动物学] 10[医学] 

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) 

主　　题：Pyrococcus furiosus(P furiosus) Uracil DNA glycosylase(UDG) Pyrococcus furiosus DNA polymeras Uracil repair in hyperthermophile 

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 ***, suggesting PfUDG in vivo functions as the re- pair enzyme to excise uracil damage in genome.