Single-molecule dynamics of the DNA-EcoRII protein complexes revealed with high-speed atomic force microscopy.
Single-molecule dynamics of the DNA-EcoRII protein complexes revealed with high-speed atomic force microscopy.
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DOI：
10.1021/bi9010368
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发表时间：
2009-11-10
期刊：
影响因子：
2.9
通讯作者：
Lyubchenko, Yuri L.
中科院分区：
文献类型：
--
作者：
Gilmore, Jamie L.;Suzuki, Yuki;Tamulaitis, Gintautas;Siksnys, Virginijus;Takeyasu, Kunio;Lyubchenko, Yuri L.
蛋白质与DNA相互作用的研究是很重要的，以获得一个基本的了解如何发生众多的生物过程，包括重组，转录，修复等，在这项研究中，我们使用的EcoRII限制性内切酶，它采用了三个位点的结合机制，以催化切割的一个单一的识别位点。使用高速原子力显微镜（HS-AFM）图像的单分子相互作用在真实的时间，我们能够观察到结合，易位，和解离机制的EcoRII蛋白。结果表明，该蛋白可以沿着DNA转位，寻找特异性结合位点。而且，一旦在单个位点特异性结合，蛋白质能够沿着DNA移位以定位第二特异性结合位点。此外，观察到EcoRII蛋白从环结构解离的两种替代模式，这导致蛋白作为单体稳定地结合到两个位点或作为二聚体结合到单个位点。从这些观察，我们提出了一个模型，在该模型中，这条途径是参与催化活性的三个网站的复合物的形成和动力学。
The study of protein interactions with DNA is important to gain a fundamental understanding of how numerous biological processes occur, including recombination, transcription, repair, etc. In this study, we use the EcoRII restriction enzyme, which employs a three-site binding mechanism in order to catalyze cleavage of a single recognition site. Using high-speed atomic force microscopy (HS-AFM) to image single-molecule interactions in real time, we were able to observe binding, translocation, and dissociation mechanisms of the EcoRII protein. The results show that the protein can translocate along DNA to search for the specific binding site. Also, once specifically bound at a single site, the protein is capable of translocating along the DNA to locate the second specific binding site. Furthermore, two alternative modes of dissociation of the EcoRII protein from the loop structure were observed, which result in the protein stably bound as monomers to two sites or bound to a single site as a dimer. From these observations, we propose a model in which this pathway is involved in the formation and dynamics of a catalytically active three-site complex.
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影响因子：
3.4
作者：
Guthold, M;Zhu, XS;Bustamante, C
通讯作者：
Bustamante, C
影响因子：
14.9
作者：
Bellamy SR;Kovacheva YS;Zulkipli IH;Halford SE
通讯作者：
Halford SE
DOI：
10.1073/pnas.211400898
发表时间：
2001-10-23
影响因子：
11.1
作者：
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通讯作者：
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影响因子：
4.6
作者：
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通讯作者：
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影响因子：
16
作者：
Mouw, Kent W.;Rowland, Sally-J.;Gajjar, Mark M.;Boocock, Martin R.;Stark, W. Marshall;Rice, Phoebe A.
通讯作者：
Rice, Phoebe A.