Synergy and antagonism in regulation of recombinant human INO80 chromatin remodeling complex

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Title: Synergy and antagonism in regulation of recombinant human INO80 chromatin remodeling complex
Authors: Willhoft, O
Bythell-Douglas, R
Mccormack, EA
Wigley, DB
Item Type: Journal Article
Abstract: We have purified a minimal core human Ino80 complex from recombinant protein expressed in insect cells. The complex comprises one subunit each of an N-terminally truncated Ino80, actin, Arp4, Arp5, Arp8, Ies2 and Ies6, together with a single heterohexamer of the Tip49a and Tip49b proteins. This core complex has nucleosome sliding activity that is similar to that of endogenous human and yeast Ino80 complexes and is also inhibited by inositol hexaphosphate (IP6). We show that IP6 is a non-competitive inhibitor that acts by blocking the stimulatory effect of nucleosomes on the ATPase activity. The IP6 binding site is located within the C-terminal region of the Ino80 subunit. We have also prepared complexes lacking combinations of Ies2 and Arp5/Ies6 subunits that reveal regulation imposed by each of them individually and synergistically that couples ATP hydrolysis to nucleosome sliding. This coupling between Ies2 and Arp5/Ies6 can be overcome in a bypass mutation of the Arp5 subunit that is active in the absence of Ies2. These studies reveal several underlying mechanisms for regulation of ATPase activity involving a complex interplay between these protein subunits and IP6 that in turn controls nucleosome sliding.
Issue Date: 2-Jun-2016
Date of Acceptance: 23-May-2016
DOI: 10.1093/nar/gkw509
ISSN: 1362-4962
Publisher: Oxford University Press
Start Page: 8179
End Page: 8188
Journal / Book Title: Nucleic Acids Research
Volume: 44
Issue: 17
Copyright Statement: © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
Sponsor/Funder: Wellcome Trust
Cancer Research UK
Funder's Grant Number: 095519/B/11/Z
Keywords: Developmental Biology
Environmental Sciences
Biological Sciences
Information And Computing Sciences
Publication Status: Published
Appears in Collections:Department of Medicine
Faculty of Medicine

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