Structural and functional characterization of deep-sea thermophilic bacteriophage GVE2 HNH endonuclease

doi:10.1038/srep42542

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	Structural and functional characterization of deep-sea thermophilic bacteriophage GVE2 HNH endonuclease
	Zhang, LK; Xu, DD; Huang, YC; Zhu, XY; Rui, MW; Wan, T; Zheng, X; Shen, YL; Chen, XD; Ma, KS; Gong, Y; Zheng X(郑欣); Gong Y(龚勇)
	2017
发表期刊	SCIENTIFIC REPORTS (IF:4.259[JCR-2016],4.847[5-Year])
ISSN	2045-2322
卷号	7 页码:42542
文章类型	Article
摘要	HNH endonucleases in bacteriophages play a variety of roles in the phage lifecycle as key components of phage DNA packaging machines. The deep-sea thermophilic bacteriophage Geobacillus virus E2 (GVE2) encodes an HNH endonuclease (GVE2 HNHE). Here, the crystal structure of GVE2 HNHE is reported. This is the first structural study of a thermostable HNH endonuclease from a thermophilic bacteriophage. Structural comparison reveals that GVE2 HNHE possesses a typical beta beta alpha-metal fold and Zn-finger motif similar to those of HNH endonucleases from other bacteriophages, apart from containing an extra a-helix, suggesting conservation of these enzymes among bacteriophages. Biochemical analysis suggests that the alanine substitutions of the conserved residues (H93, N109 and H118) in the HNH motif of GVE2 HNHE abolished 94%, 60% and 83% of nicking activity, respectively. Compared to the wild type enzyme, the H93A mutant displayed almost the same conformation while the N108A and H118A mutants had different conformations. In addition, the wild type enzyme was more thermostable than the mutants. In the presence of Mn2+ or Zn2+, the wild type enzyme displayed distinct DNA nicking patterns. However, high Mn2+ concentrations were needed for the N109A and H118A mutants to nick DNA while Zn2+ inactivated their nicking activity.
DOI	10.1038/srep42542
关键词[WOS]	RESTRICTION-ENDONUCLEASE ; DNA-BINDING ; CRYSTAL-STRUCTURE ; COLICIN E7 ; NUCLEASE DOMAIN ; CLEAVAGE ; MECHANISM ; COMPLEX ; PROTEIN ; SITE
收录类别	SCI ; PUBMED ; MEDLINE ; SCOPUS ; ADS
语种	英语
WOS研究方向	Science & Technology - Other Topics
WOS类目	Multidisciplinary Sciences
WOS记录号	WOS:000393779500001
ADS Bibcode	2017NatSR...742542Z
ADS URL	https://ui.adsabs.harvard.edu/abs/2017NatSR...742542Z
ADS引文	https://ui.adsabs.harvard.edu/abs/2017NatSR...742542Z/citations
Medline编号	MEDLINE:28211904
引用统计	正在获取... 被引频次：6 [ADS]
文献类型	期刊论文
条目标识符	https://ir.ihep.ac.cn/handle/311005/284869
专题	多学科研究中心
作者单位	中国科学院高能物理研究所
第一作者单位	中国科学院高能物理研究所
推荐引用方式 GB/T 7714	Zhang, LK,Xu, DD,Huang, YC,et al. Structural and functional characterization of deep-sea thermophilic bacteriophage GVE2 HNH endonuclease[J]. SCIENTIFIC REPORTS,2017,7:42542.
APA	Zhang, LK.,Xu, DD.,Huang, YC.,Zhu, XY.,Rui, MW.,...&龚勇.(2017).Structural and functional characterization of deep-sea thermophilic bacteriophage GVE2 HNH endonuclease.SCIENTIFIC REPORTS,7,42542.
MLA	Zhang, LK,et al."Structural and functional characterization of deep-sea thermophilic bacteriophage GVE2 HNH endonuclease".SCIENTIFIC REPORTS 7(2017):42542.