Evolutionary Conservation of Structural and Functional Coupling between the BRM AT-Hook and Bromodomain.
Evolutionary Conservation of Structural and Functional Coupling between the BRM AT-Hook and Bromodomain.
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DOI：
10.1016/j.jmb.2021.166845
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发表时间：
2021-07-09
影响因子：
5.6
通讯作者：
Musselman CA
中科院分区：
文献类型：
--
作者：
Lupo BE;Chu P;Harms MJ;Morrison EA;Musselman CA
BAF 染色质重塑复合物对于基因组调控至关重要。 BAF 的中心 ATP 酶是 BRM 或 BRG1，两者都含有 C 端溴结构域，已知与乙酰化赖氨酸相关。我们最近证明，除了乙酰赖氨酸结合之外，BRG1/BRM 溴结构域还可以通过与乙酰赖氨酸结合口袋相邻的富含赖氨酸/精氨酸的斑块与 DNA 结合。布罗莫结构域的侧翼是一个 AT 钩，由一个短的、富含脯氨酸的连接子隔开。我们之前发现 AT-hook 和 bromodomain 可以以多价方式与 DNA 结合。在这里，我们研究了这个复合模块的保守性，发现 AT 钩、连接子和富含赖氨酸/精氨酸的溴结构域补丁是古老的，保守了约 10 亿年。我们利用广泛的诱变、NMR 光谱和荧光各向异性来剖析每个保守元件对该模块与 DNA 的关联的贡献。我们的结果揭示了由连接子介导的 AT 钩和溴结构域的结构和功能耦合。布罗莫结构域上富含赖氨酸/精氨酸的片段和 AT 钩的保守元件对于 DNA 的强大亲和力至关重要，而接头的保守元件对于整体 DNA 亲和力来说是可有可无的，但对于维持 AT 钩和布罗莫结构域与 DNA 结合的相对构象至关重要。这支持 AT-hook 和溴结构域的耦合作用对于 BAF 活性很重要。
The BAF chromatin remodeling complex is critical for genome regulation. The central ATPase of BAF is either BRM or BRG1, both of which contain a C-terminal bromodomain, known to associate with acetylated lysines. We have recently demonstrated that in addition to acetyl-lysine binding, the BRG1/BRM bromodomain can associate with DNA through a lysine/arginine rich patch that is adjacent to the acetyl-lysine binding pocket. Flanking the bromodomain is an AT-hook separated by a short, proline-rich linker. We previously found that the AT-hook and bromodomain can associate with DNA in a multivalent manner. Here, we investigate the conservation of this composite module and find that the AT-hook, linker, and lysine/arginine rich bromodomain patch are ancient, conserved over ~1 billion years. We utilize extensive mutagenesis, NMR spectroscopy, and fluorescence anisotropy to dissect the contribution of each of these conserved elements in association of this module with DNA. Our results reveal a structural and functional coupling of the AT-hook and bromodomain mediated by the linker. The lysine/arginine rich patch on the bromodomain and the conserved elements of the AT-hook are critical for robust affinity for DNA, while the conserved elements of the linker are dispensable for overall DNA affinity but critical for maintaining the relative conformation of the AT-hook and bromodomain in binding to DNA. This supports that the coupled action of the AT-hook and bromodomain are important for BAF activity.
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影响因子：
7.3
作者：
Gao J;Aksoy BA;Dogrusoz U;Dresdner G;Gross B;Sumer SO;Sun Y;Jacobsen A;Sinha R;Larsson E;Cerami E;Sander C;Schultz N
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影响因子：
30.8
作者：
Kadoch, Cigall;Hargreaves, Diana C.;Hodges, Courtney;Elias, Laura;Ho, Lena;Ranish, Jeff;Crabtree, Gerald R.
通讯作者：
Crabtree, Gerald R.
影响因子：
6.8
作者：
Blessing, Charlotte;Knobloch, Gunnar;Ladurner, Andreas G.
通讯作者：
Ladurner, Andreas G.
DOI：
10.1038/nrg3413
发表时间：
2013-05
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
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