Global analysis of mutual interaction surfaces of nucleosomes with comprehensive point mutants
Global analysis of mutual interaction surfaces of nucleosomes with comprehensive point mutants
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DOI：
10.1111/j.1365-2443.2009.01350.x
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发表时间：
2009-11
期刊：
影响因子：
2.1
通讯作者：
M. Sakamoto;Shuhei Noguchi;S. Kawashima;Yusuke Okada;T. Enomoto;M. Seki;M. Horikoshi
中科院分区：
文献类型：
--
作者：
M. Sakamoto;Shuhei Noguchi;S. Kawashima;Yusuke Okada;T. Enomoto;M. Seki;M. Horikoshi
核小体中核心组蛋白的表面暴露于因子识别所需，或隐藏于组蛋白- dna和组蛋白-组蛋白相互作用中。为了了解核小体结构和功能在DNA介导的反应中协调改变的机制，有必要确定暴露和掩埋残基的作用及其功能关系。为此，我们开发了GLASP （GLobal Analysis of Surfaces by Point mutation）和GLAMP （GLobal Analysis of Mutual interaction Surfaces of multi - subunit protein complex by Point mutation）策略，这两种策略分别通过点突变对核小体中暴露的残基和埋藏的残基进行了综合分析。通过Ty抑制（Spt -表型）和对6 -杜金酸（6AU）、羟基脲（HU）和甲磺酸甲酯（MMS）的敏感性评估，四种不同的DNA介导反应需要共同和不同的GLAMP残基。组蛋白H2A和H2B界面突变的GLAMP残基主要影响Spt -表型，而不影响HU和MMS敏感性。有趣的是，在组蛋白H3-H3 '界面周围的突变GLAMP残基中，一些对Spt -表型、HU和MMS敏感性有相同的影响，而另一些对Spt -表型、HU和MMS敏感性有不同的影响。基于这些和其他结果，染色质因子与GLASP和GLAMP残基之间的功能关系为DNA介导反应中的核小体拆卸/组装过程提供了见解。
The surfaces of core histones in nucleosome are exposed as required for factor recognition, or buried for histone–DNA and histone–histone interactions. To understand the mechanisms by which nucleosome structure and function are coordinately altered in DNA‐mediated reactions, it is essential to define the roles of both exposed and buried residues and their functional relationships. For this purpose, we developed GLASP (GLobal Analysis of Surfaces by Point mutation) and GLAMP (GLobal Analysis of Mutual interaction surfaces of multi‐subunit protein complex by Point mutation) strategies, both of which are comprehensive analyses by point mutagenesis of exposed and buried residues in nucleosome, respectively. Four distinct DNA‐mediated reactions evaluated by Ty suppression (the Spt− phenotype), and sensitivities to 6‐azauracil (6AU), hydroxyurea (HU), and methyl methanesulfonate (MMS), require common and different GLAMP residues. Mutated GLAMP residues at the interface between histones H2A and H2B mainly affect the Spt− phenotype but not HU and MMS sensitivities. Interestingly, among the mutated GLAMP residues surrounding the histone H3–H3′ interface, some equally affect the Spt− phenotype, and HU and MMS sensitivities, whereas others differentially affect the Spt− phenotype, and HU and MMS sensitivities. Based on these and other results, the functional relationships among chromatin factors and GLASP and GLAMP residues provide insights into nucleosome disassembly/assembly processes in DNA‐mediated reactions.