DNA与核受体相互作用的研究:最常用方法的综述

Juliana Fattori, N. D. C. Indolfo, Jéssica L. O. Campos, N. B. Videira, A. Bridi, T. R. Doratioto, M. Assis, A. Figueira
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引用次数: 2

摘要

核受体(NRs)包括一个由配体调节的蛋白质超家族,该配体调节靶基因的表达。这些蛋白共享一个多结构域结构,包含一个n端结构域、一个高度保守的DNA结合结构域和一个配体结合结构域,具有依赖配体的激活功能。它们在发育、代谢和生理中起着关键作用,与疾病密切相关。关于这个超家族的大部分知识来自对新配体的研究,并且主要集中在配体结合域。然而,需要更多的研究来关注DNA与DNA结合域之间的相互作用,以揭示rna参与转录机制和特定基因网络的重要作用。在这里,我们的目标是讨论NRs-DNA相互作用的一些细微差别,描述这类研究中最常用的技术的细节,如凝胶移位(EMSA), DNA足迹,报告基因测定,ChIP-Seq, 3C和荧光各向异性。此外,我们的目标是提供工具,介绍这些常用方法的优缺点,在选择最适合的方法来研究rna - dna相互作用以回答特定问题时。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Investigation of Interactions between DNA and Nuclear Receptors: A Review of the Most Used Methods
Nuclear receptors (NRs) comprise a superfamily of proteins modulated by ligands that regulate the expression of target genes. These proteins share a multidomain structure harboring an N-terminal domain, a highly conserved DNA binding domain, and a ligand binding domain, which has ligand dependent activation function. They play key roles in development, metabolism, and physiology being closely related to diseases. Most of the knowledge about this superfamily emerges from investigations on new ligands and are mostly centered in the ligand binding domain. However, more investigation focusing on interactions between DNA and DNA binding domain is necessary to shed light on important roles of NRs' participation in transcriptional mechanisms and in specific genes network. Here, our goal is to discuss some nuances of NRs-DNA interaction, describing details of the most used techniques in this sort of study, such as gel shift (EMSA), DNA footprinting, reporter gene assay, ChIP-Seq, 3C, and fluorescence anisotropy. Additionally, we aim to provide tools, presenting advantages and disadvantages of these common methods, when choosing the most suitable one to study NRs-DNA interactions to answer specific questions.
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