MetJ repressor interactions with DNA probed by in-cell NMR

被引:35
作者
Augustus, Anne M. [1 ]
Reardon, Patrick N. [1 ]
Spicer, Leonard D. [1 ,2 ]
机构
[1] Duke Univ, Med Ctr, Dept Biochem, Durham, NC 27710 USA
[2] Duke Univ, Med Ctr, Dept Radiol, Durham, NC 27710 USA
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
DNA-protein interactions; gene regulation; met repressor; methionine regulon; nonspecific DNA; COLI METHIONINE REPRESSOR; ONE-DIMENSIONAL DIFFUSION; ESCHERICHIA-COLI; S-ADENOSYLMETHIONINE; COGNATE SITES; LIVING CELL; PROTEIN; SPECTROSCOPY; BINDING; GENE;
D O I
10.1073/pnas.0811130106
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];
摘要
Atomic level characterization of proteins and other macromolecules in the living cell is challenging. Recent advances in NMR instrumentation and methods, however, have enabled in-cell studies with prospects for multidimensional spectral characterization of individual macromolecular components. We present NMR data on the in-cell behavior of the MetJ repressor from Escherichia coli, a protein that regulates the expression of genes involved in methionine biosynthesis. NMR studies of whole cells along with corresponding studies in cell lysates and in vitro preparations of the pure protein give clear evidence for extensive nonspecific interactions with genomic DNA. These interactions can provide an efficient mechanism for searching out target sequences by reducing the dependence on 3-dimensional diffusion through the crowded cellular environment. DNA provides the track for MetJ to negotiate the obstacles inherent in cells and facilitates locating and binding specific repression sites, allowing for timely control of methionine biosynthesis.
引用
收藏
页码:5065 / 5069
页数:5
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