Expression, purification and characterization of the human MTA2-RBBP7 complex
Research output: Contribution to journal › Journal article › Research › peer-review
Standard
Expression, purification and characterization of the human MTA2-RBBP7 complex. / Brasen, Christoffer; Dorosz, Jerzy; Wiuf, Anders; Boesen, Thomas; Mirza, Osman; Gajhede, Michael.
In: Biochimica et biophysica acta, Vol. 1865, No. 5, 04.02.2017, p. 531-538.Research output: Contribution to journal › Journal article › Research › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Expression, purification and characterization of the human MTA2-RBBP7 complex
AU - Brasen, Christoffer
AU - Dorosz, Jerzy
AU - Wiuf, Anders
AU - Boesen, Thomas
AU - Mirza, Osman
AU - Gajhede, Michael
N1 - Copyright © 2017 Elsevier B.V. All rights reserved.
PY - 2017/2/4
Y1 - 2017/2/4
N2 - 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, p66α/β 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 MTA1-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.
AB - 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, p66α/β 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 MTA1-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.
KW - Journal Article
U2 - 10.1016/j.bbapap.2017.02.002
DO - 10.1016/j.bbapap.2017.02.002
M3 - Journal article
C2 - 28179136
VL - 1865
SP - 531
EP - 538
JO - B B A - General Subjects
JF - B B A - General Subjects
SN - 0304-4165
IS - 5
ER -
ID: 174424394