Dissecting chromatin interactions in living cells from protein mobility maps.

Fabian Erdel, Katharina Müller-Ott, Michael Baum, Malte Wachsmuth, Karsten Rippe
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引用次数: 40

Abstract

The genome of eukaryotes is organized into a dynamic nucleoprotein complex referred to as chromatin, which can adopt different functional states. Both the DNA and the protein component of chromatin are subject to various post-translational modifications that define the cell's gene expression program. Their readout and establishment occurs in a spatio-temporally coordinated manner that is controlled by numerous chromatin-interacting proteins. Binding to chromatin in living cells can be measured by a spatially resolved analysis of protein mobility using fluorescence microscopy based approaches. Recent advancements in the acquisition of protein mobility data using fluorescence bleaching and correlation methods provide data sets on diffusion coefficients, binding kinetics, and cellular concentrations on different time and length scales. The combination of different techniques is needed to dissect the complex interplay of diffusive translocations, binding events, and mobility constraints of the chromatin environment. While bleaching techniques have their strength in the characterization of particles that are immobile on the second/minute time scale, a correlation analysis is advantageous to characterize transient binding events with millisecond residence time. The application and synergy effects of the different approaches to obtain protein mobility and interaction maps in the nucleus are illustrated for the analysis of heterochromatin protein 1.

从蛋白质迁移图剖析活细胞中的染色质相互作用。
真核生物的基因组被组织成一个动态的核蛋白复合体,称为染色质,它可以采用不同的功能状态。DNA和染色质的蛋白质成分都受到各种翻译后修饰的影响,这些修饰定义了细胞的基因表达程序。它们的读取和建立以一种时空协调的方式发生,由许多染色质相互作用蛋白控制。结合染色质在活细胞中可以通过使用基于荧光显微镜的方法的蛋白质流动性的空间分辨分析来测量。利用荧光漂白和相关方法获取蛋白质迁移数据的最新进展提供了不同时间和长度尺度上的扩散系数、结合动力学和细胞浓度的数据集。需要结合不同的技术来剖析弥漫性易位、结合事件和染色质环境的迁移限制的复杂相互作用。虽然漂白技术在表征秒/分钟时间尺度上不移动的粒子方面有其优势,但相关性分析有利于表征毫秒停留时间的瞬态结合事件。在异染色质蛋白1的分析中,说明了获得蛋白质迁移和相互作用图的不同方法的应用和协同效应。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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