Iron inhibits Escherichia coli topoisomerase I activity by targeting the first two zinc-binding sites in the C-terminal domain

doi:10.1002/pro.2542

科研成果详情

题名	Iron inhibits Escherichia coli topoisomerase I activity by targeting the first two zinc-binding sites in the C-terminal domain
作者	Wang, Wu1; Su, Xiaolu1; Wang, Xiaobing2; Yang, Juanjuan1; Zhang, Ting 1; Wang, Maofeng3; Wan, Rugen 3; Tan, Guoqiang1; Lu, Jianxin 1
发表日期	2014-11
发表期刊	PROTEIN SCIENCE 影响因子和分区
语种	英语
原始文献类型	Article
关键词	Escherichia coli topoisomerase I iron binding zinc-binding motif topoisomerase activity
其他关键词	TOXIC METAL-IONS ; DNA TOPOISOMERASE ; SENSITIVITY ; MECHANISM ; PROTEINS ; CLEAVAGE ; DELETION ; RELEASE
摘要	Escherichia coli DNA topoisomerase I (TopA) contains a 67 kDa N-terminal catalytic domain and a 30 kDa C-terminal zinc-binding region (ZD domain) which has three adjacent tetra-cysteine zinc-binding motifs. Previous studies have shown that E. coli TopA can bind both iron and zinc, and that iron binding in TopA results in failure to unwind the negatively supercoiled DNA. Here, we report that each E. coli TopA monomer binds one atom of iron via the first two zinc-binding motifs in ZD domain and both the first and second zinc-binding motifs are required for iron binding in TopA. The site-directed mutagenesis studies further reveal that while the mutation of the third zinc-binding motif has very little effect on TopA's activity, mutation of the first two zinc-binding motifs in TopA greatly diminishes the topoisomerase activity in vitro and in vivo, indicating that the first two zinc-binding motifs in TopA are crucial for its function. The DNA-binding activity assay and intrinsic tryptophan fluorescence measurements show that iron binding in TopA may decrease the single-stranded (ss) DNA-binding activity of ZD domain and also change the protein structure of TopA, which subsequently modulate topoisomerase activity.
资助项目	Chinese National Natural Science Foundation Grants [31228006, 31100576]; Natural Science Foundation of Zhejiang Province Grants [LY13C050003, LY12C05003, LY13B070012]
出版者	WILEY-BLACKWELL
出版地	HOBOKEN
ISSN	0961-8368
EISSN	1469-896X
卷号	23 期号:11 页码:1619-1628
DOI	10.1002/pro.2542
页数	10
WOS类目	Biochemistry & Molecular Biology
WOS研究方向	Biochemistry & Molecular Biology
WOS记录号	WOS:000344325900013
收录类别	SCIE ; PUBMED ; SCOPUS
URL	查看原文
PubMed ID	25176012
PMC记录号	PMC4241112
SCOPUSEID	2-s2.0-84928954168
通讯作者地址	[Lu, Jianxin]Key Laboratory of Laboratory Medicine, Ministry of Education of China, Wenzhou Medical University,Wenzhou, Zhejiang,325035,China
Scopus学科分类	Biochemistry;Molecular Biology
引用统计	被引频次：1[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/15113
专题	检验医学院（生命科学学院、生物学实验教学中心）附属第一医院其他_附属东阳医院（东阳市人民医院）
通讯作者	Lu, Jianxin
作者单位	1.Key Laboratory of Laboratory Medicine, Ministry of Education of China, Wenzhou Medical University,Wenzhou, Zhejiang,325035,China; 2.Department of Rheumatism, First Affiliated Hospital of Wenzhou Medical University,Wenzhou, Zhejiang,325035,China; 3.Department of Laboratory Medicine, Affiliated Dongyang Hospital of Wenzhou Medical University,Dongyang, Zhejiang,322100,China
第一作者单位	温州医科大学
通讯作者单位	温州医科大学
第一作者的第一单位	温州医科大学
推荐引用方式 GB/T 7714	Wang, Wu,Su, Xiaolu,Wang, Xiaobing,et al. Iron inhibits Escherichia coli topoisomerase I activity by targeting the first two zinc-binding sites in the C-terminal domain[J]. PROTEIN SCIENCE,2014,23(11):1619-1628.
APA	Wang, Wu., Su, Xiaolu., Wang, Xiaobing., Yang, Juanjuan., Zhang, Ting., ... & Lu, Jianxin. (2014). Iron inhibits Escherichia coli topoisomerase I activity by targeting the first two zinc-binding sites in the C-terminal domain. PROTEIN SCIENCE, 23(11), 1619-1628.
MLA	Wang, Wu,et al."Iron inhibits Escherichia coli topoisomerase I activity by targeting the first two zinc-binding sites in the C-terminal domain".PROTEIN SCIENCE 23.11(2014):1619-1628.