用于体外功能测定的修饰染色质模板的重构。
Reconstitution of modified chromatin templates for in vitro functional assays.
作者信息
Yun Miyong, Ruan Chun, Huh Jae-Wan, Li Bing
机构信息
Department of Molecular Biology, UT Southwestern Medical Center, Dallas, TX, USA.
出版信息
Methods Mol Biol. 2012;833:237-53. doi: 10.1007/978-1-61779-477-3_15.
Abstract
To study the functions of histone modifications in the context of chromatin, it is necessary to be able to prepare nucleosomal templates that carry specific posttranslational modifications in a defined biochemical system. Here, we describe two sets of protocols for reconstituting designer nucleosomes that contain specifically modified histones. The resulting nucleosomes are suitable for electromobility shift assays, chromatin remodeling assays, and other functional and structural studies.
摘要
为了在染色质环境中研究组蛋白修饰的功能，必须能够在特定的生化系统中制备携带特定翻译后修饰的核小体模板。在此，我们描述了两组用于重构含有特定修饰组蛋白的定制核小体的方案。所得核小体适用于电泳迁移率变动分析、染色质重塑分析以及其他功能和结构研究。
相似文献
1
Reconstitution of modified chromatin templates for in vitro functional assays.用于体外功能测定的修饰染色质模板的重构。
Methods Mol Biol. 2012;833:237-53. doi: 10.1007/978-1-61779-477-3_15.
2
In Vitro Assembly of Nucleosomes for Binding/Remodeling Assays.用于结合/重塑分析的核小体体外组装
Methods Mol Biol. 2017;1528:1-17. doi: 10.1007/978-1-4939-6630-1_1.
3
Histone H4-K16 acetylation controls chromatin structure and protein interactions.组蛋白H4赖氨酸16位乙酰化调控染色质结构和蛋白质相互作用。
Science. 2006 Feb 10;311(5762):844-7. doi: 10.1126/science.1124000.
4
Histone fold modifications control nucleosome unwrapping and disassembly.组蛋白折叠修饰控制核小体的展开和拆卸。
Proc Natl Acad Sci U S A. 2011 Aug 2;108(31):12711-6. doi: 10.1073/pnas.1106264108. Epub 2011 Jul 18.
5
A defined in vitro system to study ATP-dependent remodeling of short chromatin fibers.一种用于研究短染色质纤维ATP依赖性重塑的体外系统。
Methods Mol Biol. 2012;833:255-70. doi: 10.1007/978-1-61779-477-3_16.
6
Histone acetylation: facts and questions.组蛋白乙酰化：事实与问题。
Chromosoma. 1994 Dec;103(7):441-9. doi: 10.1007/BF00337382.
7
Semisynthesis of site-specifically succinylated histone reveals that succinylation regulates nucleosome unwrapping rate and DNA accessibility.定点琥珀酰化组蛋白的半合成揭示了琥珀酰化调节核小体解缠绕速率和 DNA 可及性。
Nucleic Acids Res. 2020 Sep 25;48(17):9538-9549. doi: 10.1093/nar/gkaa663.
8
9
10
Interactions of Isw2 chromatin remodeling complex with nucleosomal arrays: analyses using recombinant yeast histones and immobilized templates.Isw2染色质重塑复合物与核小体阵列的相互作用：使用重组酵母组蛋白和固定化模板的分析
Mol Cell Biol. 2001 Mar;21(6):2098-106. doi: 10.1128/MCB.21.6.2098-2106.2001.
引用本文的文献
1
In Vitro Assembly of Nucleosomes for Binding/Remodeling Assays.用于结合/重塑分析的核小体体外组装
Methods Mol Biol. 2025;2919:1-18. doi: 10.1007/978-1-0716-4486-7_1.
2
Pioneer factors and their in vitro identification methods.先驱因子及其体外鉴定方法。
Mol Genet Genomics. 2020 Jul;295(4):825-835. doi: 10.1007/s00438-020-01675-9. Epub 2020 Apr 15.
3
Automethylation of PRC2 promotes H3K27 methylation and is impaired in H3K27M pediatric glioma.PRC2 的自动甲基化促进了 H3K27 的甲基化，并在 H3K27M 型小儿神经胶质瘤中受损。
Genes Dev. 2019 Oct 1;33(19-20):1428-1440. doi: 10.1101/gad.328773.119. Epub 2019 Sep 5.
4
Capturing the Onset of PRC2-Mediated Repressive Domain Formation.捕获 PRC2 介导的抑制性结构域形成的起始。
Mol Cell. 2018 Jun 21;70(6):1149-1162.e5. doi: 10.1016/j.molcel.2018.05.023.
5
Allosteric Activation Dictates PRC2 Activity Independent of Its Recruitment to Chromatin.变构激活决定 PRC2 活性，而不依赖于其募集到染色质上。
Mol Cell. 2018 May 3;70(3):422-434.e6. doi: 10.1016/j.molcel.2018.03.020. Epub 2018 Apr 19.
6
Identification of H3K4me1-associated proteins at mammalian enhancers.鉴定哺乳动物增强子中 H3K4me1 相关蛋白。
Nat Genet. 2018 Jan;50(1):73-82. doi: 10.1038/s41588-017-0015-6. Epub 2017 Dec 18.
7
Combinatorial Histone Readout by the Dual Plant Homeodomain (PHD) Fingers of Rco1 Mediates Rpd3S Chromatin Recruitment and the Maintenance of Transcriptional Fidelity.Rco1的双植物同源结构域（PHD）指通过组合式组蛋白识别介导Rpd3S染色质募集及转录保真度维持
J Biol Chem. 2016 Jul 8;291(28):14796-802. doi: 10.1074/jbc.M116.720193. Epub 2016 May 16.
8
Touch, act and go: landing and operating on nucleosomes.触碰、行动与前行：在核小体上着陆并进行操作。
EMBO J. 2016 Feb 15;35(4):376-88. doi: 10.15252/embj.201593377. Epub 2016 Jan 19.
9
Homodimeric PHD Domain-containing Rco1 Subunit Constitutes a Critical Interaction Hub within the Rpd3S Histone Deacetylase Complex.含同源二聚体PHD结构域的Rco1亚基构成Rpd3S组蛋白去乙酰化酶复合物中的关键相互作用中心。
J Biol Chem. 2016 Mar 4;291(10):5428-38. doi: 10.1074/jbc.M115.703637. Epub 2016 Jan 8.
10
Preparation and analysis of positioned mononucleosomes.定位单核小体的制备与分析
Methods Mol Biol. 2015;1288:15-26. doi: 10.1007/978-1-4939-2474-5_2.
本文引用的文献
1
A method for genetically installing site-specific acetylation in recombinant histones defines the effects of H3 K56 acetylation.一种在重组组蛋白中基因安装位点特异性乙酰化的方法确定了H3 K56乙酰化的作用。
Mol Cell. 2009 Oct 9;36(1):153-63. doi: 10.1016/j.molcel.2009.07.027.
2
Combined action of PHD and chromo domains directs the Rpd3S HDAC to transcribed chromatin.PHD结构域和染色质结构域的联合作用将Rpd3S组蛋白去乙酰化酶导向转录染色质。
Science. 2007 May 18;316(5827):1050-4. doi: 10.1126/science.1139004.
3
The site-specific installation of methyl-lysine analogs into recombinant histones.将甲基赖氨酸类似物位点特异性安装到重组组蛋白中。
Cell. 2007 Mar 9;128(5):1003-12. doi: 10.1016/j.cell.2006.12.041.
4
The role of chromatin during transcription.染色质在转录过程中的作用。
Cell. 2007 Feb 23;128(4):707-19. doi: 10.1016/j.cell.2007.01.015.
5
Chromatin modifications and their function.染色质修饰及其功能。
Cell. 2007 Feb 23;128(4):693-705. doi: 10.1016/j.cell.2007.02.005.
6
Histone H4-K16 acetylation controls chromatin structure and protein interactions.组蛋白H4赖氨酸16位乙酰化调控染色质结构和蛋白质相互作用。
Science. 2006 Feb 10;311(5762):844-7. doi: 10.1126/science.1124000.
7
Stable incorporation of sequence specific repressors Ash1 and Ume6 into the Rpd3L complex.序列特异性阻遏物Ash1和Ume6稳定整合到Rpd3L复合物中。
Biochim Biophys Acta. 2005 Nov 10;1731(2):77-87; discussion 75-6. doi: 10.1016/j.bbaexp.2005.09.005. Epub 2005 Oct 24.
8
Epigenetics, histone H3 variants, and the inheritance of chromatin states.表观遗传学、组蛋白H3变体与染色质状态的遗传
Cold Spring Harb Symp Quant Biol. 2004;69:235-43. doi: 10.1101/sqb.2004.69.235.
9
Reconstitution of nucleosome core particles from recombinant histones and DNA.利用重组组蛋白和DNA重建核小体核心颗粒。
Methods Enzymol. 2004;375:23-44. doi: 10.1016/s0076-6879(03)75002-2.
10
Translating the histone code.解读组蛋白密码。
Science. 2001 Aug 10;293(5532):1074-80. doi: 10.1126/science.1063127.
Zotero 插件