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Updated: Jun 23, 2025

Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis

Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis

Published on: May 17, 2016

表观遗传基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基因基

Yi-Yun Lin¹, Peter Müller¹, Evdoxia Karagianni²

¹Department of Physics and Center for NanoScience (CeNS), LMU Munich, Amaliensstrasse 54, 80799 Munich, Germany; Soft Condensed Matter and Biophysics, Department of Physics and Debye Institute for Nanomaterials Science, Utrecht University, Princetonplein 1, 3584 CC Utrecht, the Netherlands.

Journal of molecular biology

|June 22, 2024

概括

像H3K36me3和H4K5/8/12/16ac这样的翻译后修改会产生更开放的色素结构. 这些修改影响核细胞相互作用和基因组可访问性,影响基因表达.

科学领域:

* 分子生物学 * 分子生物学
* 表观遗传学是一种表观遗传学.
* 生物物理生物物理

背景情况:

*核细胞是控制基因组可访问性的基本染色体单元.
* 基因组上的翻译后修饰 (PTM) 动态调节染色质结构和基因表达.
*了解PTM对核细胞相互作用的影响对于破译基因调节至关重要.

研究的目的:

* 为了研究H3K36me3和H4K5/8/12/16ac对三核细胞组的结构和动态影响.
*阐明这些PTM如何影响核体-核体相互作用和整体染色体构造.
* 建立一种定量单分子方法来研究染色质中的PTM.

主要方法:

* 原子力显微镜 (AFM) 用于直接可视化核体构造.
*磁性子强迫光谱法测量DNA包裹和核细胞相互作用.
*利用三核子组为量化分析的可处理模型系统.

主要成果:

* H3K36me3和H4K5/8/12/16ac修改后的核细胞与未经修改的核细胞相比,具有更开放,更宽松的形状.
* PTMs减少了DNA的外转包裹,并减少了核体与核体之间的相互作用.
* H4K5/8/12/16ac驱动的阵列开放主要是由于减少了核体间相互作用,而不是单个核体动力学.

结论:

* H3K36me3和H4K5/8/12/16ac PTM显著改变了核细胞组的结构和动态.
*核体与核体之间的相互作用在高阶染色体结构中起着关键作用,可能会超越个体核体的呼吸动态.
* 开发的方法适用于研究其他PTM和核细胞重塑剂.

关键词:

通过乙化处理.在这种情况下,染色染色素甲基化处理甲基化处理核细胞是核细胞的组成部分.单分子方法的方法.

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Isolation and Cultivation of Neural Progenitors Followed by Chromatin-Immunoprecipitation of Histone 3 Lysine 79 Dimethylation Mark

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Purification of H3 and H4 Histone Proteins and the Quantification of Acetylated Histone Marks in Cells and Brain Tissue

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