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首页> 外文期刊>EMBO Journal >The McrBC restriction endonuclease assembles into a ring structure in the presence of G nucleotides.
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The McrBC restriction endonuclease assembles into a ring structure in the presence of G nucleotides.

机译：的McrBC限制性内切核酸酶组装G环结构的存在核苷酸。

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摘要

McrBC from Escherichia coli K-12 is a restriction enzyme that belongs to the family of AAA(+) proteins and cuts DNA containing modified cytosines. Two proteins are expressed from the mcrB gene: a full-length version, McrB(L), and a short version, McrB(S). McrB(L) binds specifically to the methylated recognition site and is, therefore, the DNA-binding moiety of the McrBC endonuclease. McrB(S) is devoid of DNA-binding activity. We observed that the quaternary structure of the endonuclease depends on binding of the cofactors. In gel filtration experiments, McrB(L) and McrB(S) form high molecular weight oligomers in the presence of Mg(2+) and GTP, GDP or GTP-gamma-S. Oligomerization did not require the presence of DNA and was independent of GTP hydrolysis. Electron micrographs of negatively stained McrB(L) and McrB(S) revealed ring-shaped particles with a central channel. Mass analysis by scanning transmission electron microscopy indicates that McrB(L) and McrB(S) form single heptameric rings as well as tetradecamers. In the presence of McrC, a subunit that is essential for DNA cleavage, the tetradecameric species was the major form of the endonuclease.
机译：从大肠杆菌McrBC k - 12是一个限制酶,属于家族AAA (+)蛋白质和DNA包含修改胞核嘧啶。mcrB基因:一个完整的版本,mcrB(左),和一个短的版本,McrB(年代)。特别是甲基化识别网站,因此,dna的一部分McrBC核酸内切酶。dna结合活性。四级结构的核酸内切酶绑定的代数余子式。实验中,McrB(左)和McrB (S)形成高分子量低聚物的存在镁(2 +)和三磷酸鸟苷、GDP或GTP-gamma-S。齐聚反应不需要的存在DNA和独立于三磷酸鸟苷水解。负染色的电子显微图McrB(左)和McrB (S)显示环形粒子与中央通道。扫描透射电子显微镜表明McrB(左)和McrB (S)形式单一heptameric环以及tetradecamers。McrC面前,亚基是至关重要的DNA乳沟,tetradecameric物种核酸内切酶的主要形式。

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