Combinatorial profiling of chromatin binding modules reveals multisite discrimination.
Combinatorial profiling of chromatin binding modules reveals multisite discrimination.
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染色质结合模块的组合分析揭示了多位点歧视。
DOI：
10.1038/nchembio.319
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发表时间：
2010-04
影响因子：
14.8
通讯作者：
Denu, John M.
中科院分区：
文献类型：
--
作者：
Garske, Adam L.;Oliver, Samuel S.;Wagner, Elise K.;Musselman, Catherine A.;LeRoy, Gary;Garcia, Benjamin A.;Kutateladze, Tatiana G.;Denu, John M.
翻译后修饰（PTMs）和染色质结合蛋白之间的特定相互作用是“组蛋白密码”概念的核心。本研究利用一个基于组蛋白H3 n端、包含5000个成员的ptm随机组合肽文库，研究了6个染色质结合模块的多位点特异性，这些模块读取了K4的甲基化状态。我们发现T3磷酸化、R2甲基化和T6磷酸化是调节组蛋白H3识别和结合能力的关键ptm。值得注意的是，T6的磷酸化对蛋白质结合的影响最为多样，这表明H3尾部读卡器有重要的调控机制。质谱分析和基于抗体的证据表明，这种先前未表征的修饰存在于天然H3上，ING2的核磁共振分析揭示了区分的结构基础。这些研究揭示了一个连续的结合亲和力，其中多位点PTM识别涉及开关和变阻器样特性，产生依赖于固有“阅读器”特异性的分级效应。
Specific interactions between post-translational modifications (PTMs) and chromatin-binding proteins are central to the idea of a ‘histone code’. Here, a 5000-member, PTM-randomized, combinatorial peptide library based on the N-terminus of histone H3 was utilized to interrogate multi-site specificity of six chromatin-binding modules, which read the methylation status of K4. We found that T3 phosphorylation, R2 methylation, and T6 phosphorylation are critical additional PTMs that modulate the ability to recognize and bind histone H3. Notably, phosphorylation of T6 yielded the most varied effect on protein binding, suggesting an important regulatory mechanism for readers of the H3 tail. Mass spectrometry and antibody-based evidence indicate that this previously uncharacterized modification exists on native H3, and NMR analysis of ING2 revealed the structural basis for discrimination. These investigations reveal a continuum of binding affinities in which multi-site PTM recognition involves both switch- and rheostat-like properties, yielding graded effects that depend on the inherent ‘reader’ specificity.
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影响因子：
64.5
作者：
Guenther, Matthew G.;Levine, Stuart S.;Young, Richard A.
通讯作者：
Young, Richard A.
影响因子：
7
作者：
Huang, Hailiang;Maertens, Alexandra M.;Bader, Joel S.
通讯作者：
Bader, Joel S.
影响因子：
56.9
作者：
Huang, Y;Fang, J;Xu, RM
通讯作者：
Xu, RM
影响因子：
7.7
作者：
Org, Tonis;Chignola, Francesca;Peterson, Paert
通讯作者：
Peterson, Paert
DOI：
10.1073/pnas.0709170104
发表时间：
2007-11-27
影响因子：
11.1
作者：
Ramon-Maiques, Santiago;Kuo, Alex J.;Yang, Wei
通讯作者：
Yang, Wei