The DNA-binding domain of yeast Rap1 interacts with double-stranded DNA in multiple binding modes.
The DNA-binding domain of yeast Rap1 interacts with double-stranded DNA in multiple binding modes.
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酵母RAP1的DNA结合结构域与多种结合模式中的双链DNA相互作用。
DOI：
10.1021/bi501049b
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发表时间：
2014-12-09
期刊：
影响因子：
2.9
通讯作者：
Galletto, Roberto
中科院分区：
文献类型：
--
作者：
Feldmann, Erik A.;Galletto, Roberto
酿酒酵母阻遏激活蛋白 1 (Rap1) 是一种参与 DNA 调控多个步骤的必需蛋白，作为转录激活剂、沉默元件的阻遏蛋白以及端粒处庇护蛋白样复合物的主要组成部分。 Rap1 的所有已知功能都需要 DNA 结合域与其识别序列之间已知的高亲和力和特异性相互作用。在这项工作中，我们重点研究 Rap1 (Rap1DBD) 的 DNA 结合域与双链 DNA 底物的相互作用。出乎意料的是，我们发现，虽然Rap1DBD与其DNA识别位点形成高亲和力1:1复合物，但它也可以与DNA上较高化学计量形成低亲和力复合物。这些较低亲和力的相互作用与识别序列的存在无关，我们认为它们源于 Rap1DBD 以两种不同的结合模式与 DNA 结合的能力。在一种高亲和力结合模式中，Rap1DBD 可能以在可用晶体结构中观察到的构象结合。在另一种低亲和力结合模式中，我们提出 Rap1DBD 的单个 Myb 样结构域与 DNA 相互作用，允许多个蛋白质分子与 DNA 底物结合。我们的研究结果表明 Rap1DBD 不仅仅将蛋白质靶向其识别序列，而且它可能是一个可能的调节点。
Saccharomyces cerevisiae repressor-activator protein 1 (Rap1) is an essential protein involved in multiple steps of DNA regulation, as an activator in transcription, as a repressor at silencer elements, and as a major component of the shelterin-like complex at telomeres. All the known functions of Rap1 require the known high-affinity and specific interaction of the DNA-binding domain with its recognition sequences. In this work, we focus on the interaction of the DNA-binding domain of Rap1 (Rap1DBD) with double-stranded DNA substrates. Unexpectedly, we found that while Rap1DBD forms a high-affinity 1:1 complex with its DNA recognition site, it can also form lower-affinity complexes with higher stoichiometries on DNA. These lower-affinity interactions are independent of the presence of the recognition sequence, and we propose they originate from the ability of Rap1DBD to bind to DNA in two different binding modes. In one high-affinity binding mode, Rap1DBD likely binds in the conformation observed in the available crystal structures. In the other alternative lower-affinity binding mode, we propose that a single Myb-like domain of the Rap1DBD makes interactions with DNA, allowing for more than one protein molecule to bind to the DNA substrates. Our findings suggest that the Rap1DBD does not simply target the protein to its recognition sequence but rather it might be a possible point of regulation.
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影响因子：
14.9
作者：
GRAHAM, IR;CHAMBERS, A
通讯作者：
CHAMBERS, A
影响因子：
5.6
作者：
Del Vescovo, V;De Sanctis, V;Negri, R
通讯作者：
Negri, R
影响因子：
2.9
作者：
Barranco-Medina, Sergio;Galletto, Roberto
通讯作者：
Galletto, Roberto
DOI：
10.1007/978-1-62703-032-8_4
发表时间：
2012-01-01
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Kozlov, Alexander G;Galletto, Roberto;Lohman, Timothy M
通讯作者：
Lohman, Timothy M
影响因子：
2.9
作者：
Kumaran, Sangaralingam;Kozlov, Alexander G.;Lohman, Timothy M.
通讯作者：
Lohman, Timothy M.