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Structural and Functional Characterization of Escherichia coli Toxin-Antitoxin Complex DinJ-YafQ
Liang YJ(梁雅静); Gao ZQ(高增强); Wang F(王斐); Liang, YJ; Gao, ZQ; Wang, F; Zhang, YL; Dong, YH; Liu, QS;; Dong YH(董宇辉); Liu QS(刘全胜)
2014
发表期刊JOURNAL OF BIOLOGICAL CHEMISTRY
卷号289期号:30页码:21191-21202
摘要Toxin YafQ functions as a ribonuclease in the dinJ-yafQ toxin-antitoxin system of Escherichia coli. Antitoxin DinJ neutralizes YafQ-mediated toxicity by forming a stable protein complex. Here, crystal structures of the (DinJ)(2)-(YafQ)(2) complex and the isolated YafQ toxin have been determined. The structure of the heterotetrameric complex (DinJ)(2)-(YafQ)(2) revealed that the N-terminal region of DinJ folds into a ribbonhelix-helix motif and dimerizes for DNA recognition, and the C-terminal portion of each DinJ exclusively wraps around a YafQ molecule. Upon incorporation into the heterotetrameric complex, a conformational change of YafQ in close proximity to the catalytic site of the typical microbial ribonuclease fold was observed and validated. Mutagenesis experiments revealed that a DinJ mutant restored YafQ RNase activity in a tetramer complex in vitro but not in vivo. An electrophoretic mobility shift assay showed that one of the palindromic sequences present in the upstream intergenic region of DinJ served as a binding sequences for both the DinJ-YafQ complex and the antitoxin DinJ alone. Based on structure-guided and site-directed mutagenesis of DinJ-YafQ, we showed that two pairs of amino acids in DinJ were important for DNA binding; the R8A and K16A substitutions and the S31A and R35A substitutions in DinJ abolished the DNA binding ability of the DinJ-YafQ complex.
学科领域Biochemistry & Molecular Biology
DOI10.1074/jbc.M114.559773
收录类别SCI
WOS记录号WOS:000339396600068
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被引频次:12[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.ihep.ac.cn/handle/311005/225067
专题多学科研究中心
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Liang YJ,Gao ZQ,Wang F,et al. Structural and Functional Characterization of Escherichia coli Toxin-Antitoxin Complex DinJ-YafQ[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2014,289(30):21191-21202.
APA 梁雅静.,高增强.,王斐.,Liang, YJ.,Gao, ZQ.,...&刘全胜.(2014).Structural and Functional Characterization of Escherichia coli Toxin-Antitoxin Complex DinJ-YafQ.JOURNAL OF BIOLOGICAL CHEMISTRY,289(30),21191-21202.
MLA 梁雅静,et al."Structural and Functional Characterization of Escherichia coli Toxin-Antitoxin Complex DinJ-YafQ".JOURNAL OF BIOLOGICAL CHEMISTRY 289.30(2014):21191-21202.
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