Torrado Mario, Low Jason K K, Silva Ana P G, Schmidberger Jason W, Sana Maryam, Sharifi Tabar Mehdi, Isilak Musa E, Winning Courtney S, Kwong Cherry, Bedward Max J, Sperlazza Mary J, Williams David C, Shepherd Nicholas E, Mackay Joel P
School of Life and Environmental Sciences, University of Sydney, Australia.
Department of Pathology and Laboratory Medicine, The University of North Carolina - Chapel Hill, NC, USA.
FEBS J. 2017 Dec;284(24):4216-4232. doi: 10.1111/febs.14301. Epub 2017 Nov 13.
The nucleosome remodelling and deacetylase (NuRD) complex is essential for the development of complex animals. NuRD has roles in regulating gene expression and repairing damaged DNA. The complex comprises at least six proteins with two or more paralogues of each protein routinely identified when the complex is purified from cell extracts. To understand the structure and function of NuRD, a map of direct subunit interactions is needed. Dozens of published studies have attempted to define direct inter-subunit connectivities. We propose that conclusions reported in many such studies are in fact ambiguous for one of several reasons. First, the expression of many NuRD subunits in bacteria is unlikely to lead to folded, active protein. Second, interaction studies carried out in cells that contain endogenous NuRD complex can lead to false positives through bridging of target proteins by endogenous components. Combining existing information on NuRD structure with a protocol designed to minimize false positives, we report a conservative and robust interaction map for the NuRD complex. We also suggest a 3D model of the complex that brings together the existing data on the complex. The issues and strategies discussed herein are also applicable to the analysis of a wide range of multi-subunit complexes.
Micrococcal nuclease (MNase), EC 3.1.31.1; histone deacetylase (HDAC), EC 3.5.1.98.
核小体重塑去乙酰化酶（NuRD）复合物对于复杂动物的发育至关重要。NuRD在调节基因表达和修复受损DNA方面发挥作用。该复合物至少由六种蛋白质组成，从细胞提取物中纯化该复合物时，通常会鉴定出每种蛋白质的两个或更多个旁系同源物。为了了解NuRD的结构和功能，需要一张直接亚基相互作用的图谱。数十项已发表的研究试图定义直接的亚基间连接性。我们认为，许多此类研究报告的结论实际上因多种原因之一而模棱两可。首先，许多NuRD亚基在细菌中的表达不太可能产生折叠的活性蛋白。其次，在含有内源性NuRD复合物的细胞中进行的相互作用研究可能会通过内源性成分桥接靶蛋白而导致假阳性。结合关于NuRD结构的现有信息以及旨在最小化假阳性的方案，我们报告了一张关于NuRD复合物的保守且可靠的相互作用图谱。我们还提出了该复合物的三维模型，该模型整合了关于该复合物的现有数据。本文讨论的问题和策略也适用于广泛的多亚基复合物的分析。
微球菌核酸酶（MNase），EC 3.1.31.1；组蛋白去乙酰化酶（HDAC），EC 3.5.1.98。
