Mechanistic Insights into Autoinhibition of the Oncogenic Chromatin Remodeler ALC1

TY - JOUR

T1 - Mechanistic Insights into Autoinhibition of the Oncogenic Chromatin Remodeler ALC1

AU - Lehmann, Laura C.

AU - Hewitt, Graeme

AU - Aibara, Shintaro

AU - Leitner, Alexander

AU - Marklund, Emil

AU - Maslen, Sarah L.

AU - Maturi, Varun

AU - Chen, Yang

AU - van der Spoel, David

AU - Skehel, J. Mark

AU - Moustakas, Aristidis

AU - Boulton, Simon J.

AU - Deindl, Sebastian

N1 - Funding Information: We thank Evalena Andersson for initial work; Greg Bowman for nucleosomes; Stefan Knight, Markus Seeliger, and Sofia Pytharopoulou for discussions; Ruedi Aebersold for access to instrumentation; Robert Rambo, Nathan Cowieson, and the staff at DLS and ESRF for generous technical support. S.D. acknowledges support from a European Research Council (ERC) Starting Grant (ChromatinRemodelling), the Swedish Research Council ( VR 2015-04568 ), the Knut and Alice Wallenberg Foundation ( KAW/WAF 2014.0183 ), and the SciLifeLab . A.L. received funding from the ETH Zurich . The Boulton Lab is supported by the Francis Crick Institute , which receives its core funding from Cancer Research UK ( FC0010048 ), the UK Medical Research Council ( FC0010048 ), and the Wellcome Trust ( FC0010048 ). S.J.B. is also funded by an ERC Advanced Investigator (TelMetab) and a Wellcome Trust Senior Investigator Grant . D.v.d.S. acknowledges a VR computer time grant ( SNIC2013-26-6 ) and S.A. a FEBS Long-Term Fellowship . Funding Information: We thank Evalena Andersson for initial work; Greg Bowman for nucleosomes; Stefan Knight, Markus Seeliger, and Sofia Pytharopoulou for discussions; Ruedi Aebersold for access to instrumentation; Robert Rambo, Nathan Cowieson, and the staff at DLS and ESRF for generous technical support. S.D. acknowledges support from a European Research Council (ERC) Starting Grant (ChromatinRemodelling), the Swedish Research Council (VR 2015-04568), the Knut and Alice Wallenberg Foundation (KAW/WAF 2014.0183), and the SciLifeLab. A.L. received funding from the ETH Zurich. The Boulton Lab is supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC0010048), the UK Medical Research Council (FC0010048), and the Wellcome Trust (FC0010048). S.J.B. is also funded by an ERC Advanced Investigator (TelMetab) and a Wellcome Trust Senior Investigator Grant. D.v.d.S. acknowledges a VR computer time grant (SNIC2013-26-6) and S.A. a FEBS Long-Term Fellowship. Publisher Copyright: © 2017 Elsevier Inc.

PY - 2017/12/7

Y1 - 2017/12/7

N2 - Human ALC1 is an oncogene-encoded chromatin-remodeling enzyme required for DNA repair that possesses a poly(ADP-ribose) (PAR)-binding macro domain. Its engagement with PARylated PARP1 activates ALC1 at sites of DNA damage, but the underlying mechanism remains unclear. Here, we establish a dual role for the macro domain in autoinhibition of ALC1 ATPase activity and coupling to nucleosome mobilization. In the absence of DNA damage, an inactive conformation of the ATPase is maintained by juxtaposition of the macro domain against predominantly the C-terminal ATPase lobe through conserved electrostatic interactions. Mutations within this interface displace the macro domain, constitutively activate the ALC1 ATPase independent of PARylated PARP1, and alter the dynamics of ALC1 recruitment at DNA damage sites. Upon DNA damage, binding of PARylated PARP1 by the macro domain induces a conformational change that relieves autoinhibitory interactions with the ATPase motor, which selectively activates ALC1 remodeling upon recruitment to sites of DNA damage. Lehmann et al. employ an integrative structural approach with biochemistry, cell-based assays, and in vivo imaging to investigate the regulation of ALC1. An unappreciated functional crosstalk between the PAR-binding macro domain and the ATPase underlies autoinhibition and activation of ALC1.

AB - Human ALC1 is an oncogene-encoded chromatin-remodeling enzyme required for DNA repair that possesses a poly(ADP-ribose) (PAR)-binding macro domain. Its engagement with PARylated PARP1 activates ALC1 at sites of DNA damage, but the underlying mechanism remains unclear. Here, we establish a dual role for the macro domain in autoinhibition of ALC1 ATPase activity and coupling to nucleosome mobilization. In the absence of DNA damage, an inactive conformation of the ATPase is maintained by juxtaposition of the macro domain against predominantly the C-terminal ATPase lobe through conserved electrostatic interactions. Mutations within this interface displace the macro domain, constitutively activate the ALC1 ATPase independent of PARylated PARP1, and alter the dynamics of ALC1 recruitment at DNA damage sites. Upon DNA damage, binding of PARylated PARP1 by the macro domain induces a conformational change that relieves autoinhibitory interactions with the ATPase motor, which selectively activates ALC1 remodeling upon recruitment to sites of DNA damage. Lehmann et al. employ an integrative structural approach with biochemistry, cell-based assays, and in vivo imaging to investigate the regulation of ALC1. An unappreciated functional crosstalk between the PAR-binding macro domain and the ATPase underlies autoinhibition and activation of ALC1.

KW - ADP-ribosylation

KW - allosteric regulation

KW - allostery

KW - ATP-dependent chromatin remodeler

KW - ATPase

KW - chromatin remodeling

KW - DNA repair

KW - macro domain

KW - PARP

KW - structure

UR - http://www.scopus.com/inward/record.url?scp=85037847030&partnerID=8YFLogxK

U2 - 10.1016/j.molcel.2017.10.017

DO - 10.1016/j.molcel.2017.10.017

M3 - Article

C2 - 29220652

AN - SCOPUS:85037847030

SN - 1097-2765

VL - 68

SP - 847-859.e7

JO - MOLECULAR CELL

JF - MOLECULAR CELL

IS - 5

ER -