Crystal structures of phage NrS-1 N300-dNTPs-Mg2+ complex provide molecular mechanisms for substrate specificity

doi:10.1016/j.bbrc.2019.05.162

CORC > 上海应用物理研究所 > 中国科学院上海应用物理研究所 > 中科院上海应用物理研究所2011-2017年

	Crystal structures of phage NrS-1 N300-dNTPs-Mg2+ complex provide molecular mechanisms for substrate specificity
	Guo, HJ ; Li, MJ ; Wu, H ; Wang, WW ; Yu, F ; He, JH
刊名	BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
	2019
卷号	515 期号:4 页码:551-557
关键词	RIBONUCLEOTIDE INCORPORATION DNA-SYNTHESIS PRIMASE OPTIMIZATION DEGRADATION PRIMPOL DOMAIN
ISSN号	0006-291X
DOI	10.1016/j.bbrc.2019.05.162
文献子类	期刊论文
英文摘要	A novel DNA polymerase from the deep-sea vent phage NrS-1, was characterized as a primase-polymerase (referred to as prim-pol), which works as a self-priming DNA polymerase to synthesize de novo long DNA strands. Functional research on the NrS-1 prim-pol illustrated that the N-terminal 300 residues (referred to as N300) have de novo synthesis activity similar to that of the full-length enzyme. Just like other prim-pols, NrS-1 prim-pol was able to initiate DNA synthesis, proficiently discriminating against ribonucleotides (NTPs), exclusively using deoxynucleotides (dNTPs). However, the structural basis for this discrimination is not well understood. Here, the three kinds of crystal structures of N300-dNTPs-Mg2+ complex were determined. These complex structures shared the identical steric architecture and hydrogen-bond interactions in the catalytic center. The results of biochemical studies indicated that R145 possibly plays an indispensable role in the primer extension. Mutagenesis and structural simulation showed that the backbone carboxyl group of Y146, as a potential sugar selector, was involved in steric clashing with the incoming 2'-OH group of NTPs. However, the mechanism of substrate discrimination probably was different from that of other prim-pols, according to the structural analyses and sequence comparison. (C) 2019 Elsevier Inc. All rights reserved.
语种	英语
内容类型	期刊论文
源URL	[http://ir.sinap.ac.cn/handle/331007/32017]
专题	上海应用物理研究所_中科院上海应用物理研究所2011-2017年
作者单位	1.Chinese Acad Sci, Shanghai Inst Appl Phys, Shanghai 201800, Peoples R China; 2.Chinese Acad Sci, Shanghai Adv Res Inst, Shanghai 201204, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China;
推荐引用方式 GB/T 7714	Guo, HJ,Li, MJ,Wu, H,et al. Crystal structures of phage NrS-1 N300-dNTPs-Mg2+ complex provide molecular mechanisms for substrate specificity[J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,2019,515(4):551-557.
APA	Guo, HJ,Li, MJ,Wu, H,Wang, WW,Yu, F,&He, JH.(2019).Crystal structures of phage NrS-1 N300-dNTPs-Mg2+ complex provide molecular mechanisms for substrate specificity.BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,515(4),551-557.
MLA	Guo, HJ,et al."Crystal structures of phage NrS-1 N300-dNTPs-Mg2+ complex provide molecular mechanisms for substrate specificity".BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 515.4(2019):551-557.