Expression, purification and characterization of the human MTA2-RBBP7 complex

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  • Christoffer Brasen, Københavns Universitet
  • ,
  • Jerzy Dorosz, Københavns Universitet
  • ,
  • Anders Wiuf, Københavns Universitet
  • ,
  • Thomas Boesen
  • Osman Mirza, Københavns Universitet
  • ,
  • Michael Gajhede, Københavns Universitet

The repressive Nucleosome Remodeling and histone Deacetylation (NuRD) complex remodels the chromatin structure by coupling ATP-dependent remodeling activity with histone deacetylase function and plays important roles in regulating gene transcription, DNA damage repair and chromatin assembly. The complex is composed of six subunits: Metastasis Associated proteins MTA1/2/3 initially recruit histone chaperones RBBP4/7 followed by the histone deacetylases HDAC1/2 forming a core complex. Further association of the CpG-binding protein MBD2/3, p66 alpha/beta and the ATP-dependent helicase CDH3/4 constitutes the NuRD complex. Recent structural studies on truncated human proteins or orthologous have revealed that the stoichiometry of the MTAl-RBBP4 complex is 2:4. This study reports expression and purification of the intact human MTA2-RBBP7 complex using HEK293F cells as expression system. In analogy with findings on the Drosophila NuRD complex, we find that also the human MTA-RBBP can be isolated in vitro. Taken together with previous findings this suggests, that MTA-RBBP is a stable complex, with a central role in the initial assembly of the human NuRD complex. Refined 3D volumes of the complex generated from negative stain electron microscopy (EM) data reveals an elongated architecture that is capable of hinge like motion around the center of the particle. (C) 2017 Elsevier B.V. All rights reserved.

Original languageEnglish
JournalB B A - Proteins and Proteomics
Volume1865
Issue5
Pages (from-to)531-538
Number of pages8
ISSN1570-9639
DOIs
Publication statusPublished - May 2017

    Research areas

  • NuRD complex, Transient HEK293F cell expression, Negative stain electron microscopy, CHROMATIN REMODELING COMPLEX, HISTONE DEACETYLASE, BAH DOMAIN, ANGSTROM RESOLUTION, ELECTRON-MICROSCOPY, CRYSTAL-STRUCTURE, STRUCTURAL BASIS, NUCLEOSOME, NURD, CANCER

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