The Nucleosome Remodeling and Deacetylase Complex NuRD Is Built from Preformed Catalytically Active Sub-modules

JOURNAL OF MOLECULAR BIOLOGY

卷:428

Article

Zhang, W.¹  Aubert, A.²  de Segura, J. M. Gomez²  Karuppasamy, M.²  Basu, S.¹  Murthy, A. S.¹  Diamante, A.¹  Drury, T. A.¹  Balmer, J.¹  Cramard, J.³  Watson, A. A.¹  Lando, D.¹  Lee, S. F.⁴  Palayret, M.⁴  Kloet, S. L.⁵  Smits, A. H.⁵  Deery, M. J.⁶  Vermeulen, M.⁵  Hendrich, B.³  Klenerman, D.⁴  Schaffitzel, C.^2,7  Berger, I.^2,7  Laue, E. D.¹ 

[1] Univ Cambridge, Dept Biochem, 80 Tennis Court Rd, Cambridge CB2 1GA, England

[2] EMBL Grenoble, CS 90181, 71 Ave Martyrs, F-38042 Grenoble 9, France

[3] Univ Cambridge, Med Res Council Stem Cell Inst, Wellcome Trust, Gleeson Bldg,Tennis Court Rd, Cambridge CB2 1QR, England

[4] Univ Cambridge, Dept Chem, Lensfield Rd, Cambridge CB2 1EW, England

[5] Radboud Inst Mol Life Sci, Dept Mol Biol, M850-3-79 Geett Grooteplein Zuid 30, NL-6525 GA Nijmegen, Netherlands

[6] Univ Cambridge, Dept Biochem, Cambridge Syst Biol Ctr, Wellcome Trust Stem Cell Bldg,Tennis Court Rd, Cambridge CB2 1QR, England

[7] Univ Walk, Univ Bristol, Sch Biochem, Clifton BS8 1TD, England

关键词: chromatin-remodeling;    complex assembly;    histone modification;    nucleosome remodeling and deacetylase NuRD;    transcription;   

DOI  :  10.1016/j.jmb.2016.04.025

来源: Elsevier

【 摘 要 】

The nucleosome remodeling deacetylase (NuRD) complex is a highly conserved regulator of chromatin structure and transcription. Structural studies have shed light on this and other chromatin modifying machines, but much less is known about how they assemble and whether stable and functional sub-modules exist that retain enzymatic activity. Purification of the endogenous Drosophila NuRD complex shows that it consists of a stable core of subunits, while others, in particular the chromatin remodeler CHD4, associate transiently. To dissect the assembly and activity of NuRD, we systematically produced all possible combinations of different components using the MultiBac system, and determined their activity and biophysical properties. We carried out single-molecule imaging of CHD4 in live mouse embryonic stem cells, in the presence and absence of one of core components (MBD3), to show how the core deacetylase and chromatin-remodeling sub-modules associate in vivo. Our experiments suggest a pathway for the assembly of NuRD via preformed and active sub-modules. These retain enzymatic activity and are present in both the nucleus and the cytosol, an outcome with important implications for understanding NuRD function. (C) 2016 The Authors. Published by Elsevier Ltd.

【 授权许可】

Free