CRCU

Chemical Research in Chinese Universities ›› 2012, Vol. 28 ›› Issue (3): 477-482.

• Articles • Previous Articles     Next Articles

Biochemical Characterization of Uracil-DNA Glycosylase from Pyrococcus furiosus

LIN Li-bo, LIU Yu-fen, LIU Xi-peng, LIU Jian-hua   

  1. Key Laboratory of Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiaotong University, Shanghai 200240, P. R. China
  • Received:2011-08-22 Revised:2011-09-19 Online:2012-05-25 Published:2012-05-03
  • Contact: LIU Xi-peng E-mail:xpliu@sjtu.edu.cn
  • Supported by:

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

PDF (PC)

Abstract: 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 excision 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 NaCl. The phosphorothioates near uracil strongly inhibit the excision of uracil by PfUDG. Interestingly, pfuDNA(Pyrococcus furiosus DNA) polymerase, which tightly binds the uracil- carrying oligonucleotide, does not inhibit the excision by PfUDG, suggesting PfUDG in vivo functions as the repair enzyme to excise uracil damage in genome.

Key words: Pyrococcus furiosus(P. furiosus), Uracil DNA glycosylase(UDG), Pyrococcus furiosus DNA polymeras, Uracil repair in hyperthermophile