著者
中村 照也 山縣 ゆり子 YANG Wei
出版者
一般社団法人 日本生物物理学会
雑誌
生物物理 (ISSN:05824052)
巻号頁・発行日
vol.53, no.5, pp.254-257, 2013 (Released:2013-09-25)
参考文献数
13
被引用文献数
1 1

A number of structural and kinetic studies of DNA polymerases have proposed the catalytic mechanism of the nucleotidyl-transfer reaction. However, the actual process has never been visualized. Here we show the nucleotidyl-transfer reaction process catalyzed by human DNA polymerase η using time-resolved protein crystallography. In sequence, the nucleophile 3′-OH is deprotonated, the deoxyribose at the primer end converts from C2′-endo to C3′-endo, and the nucleophile and the α-phosphate of dATP approach each other to form the new bond. A third Mg2+ ion, which arrives with the new bond and stabilizes the intermediate state, may be an unappreciated feature of the two-metal-ion mechanism.
著者
中村 照也
出版者
日本結晶学会
雑誌
日本結晶学会誌 (ISSN:03694585)
巻号頁・発行日
vol.56, no.2, pp.123-128, 2014-04-30 (Released:2014-05-01)
参考文献数
30

DNA, which carries genetic information, is easily damaged by reactive oxygen species,UV radiation and chemical agents, and DNA integrity is maintained by various nucleic acid enzymes. We carried out X-ray crystallographic studies of two types of nucleic acid enzymes, oxidative nucleotide hydrolase MutT and translesion DNA polymerase η. In this review, I will describe the mechanism of high substrate specificity for oxidative nucleotides by E. coli MutT and the visualization of nucleotidyl transfer reaction by human DNA polymerase η.