@article{fd3aa1f3c63845459eff5336966256b7,
title = "Structure and dynamics of the chromatin remodeler alc1 bound to a parylated nucleosome",
abstract = "The chromatin remodeler ALC1 is recruited to and activated by DNA damage-induced poly(ADP-ribose) (PAR) chains deposited by PARP1/PARP2/HPF1 upon detection of DNA lesions. ALC1 has emerged as a candidate drug target for cancer therapy as its loss confers synthetic lethality in homologous recombination-deficient cells. However, structure-based drug design and molecular analysis of ALC1 have been hindered by the requirement for PARylation and the highly heterogeneous nature of this post-translational modification. Here, we reconstituted an ALC1 and PARylated nucleosome complex modified in vitro using PARP2 and HPF1. This complex was amenable to cryo-EM structure determination without cross-linking, which enabled visualization of several intermediate states of ALC1 from the recognition of the PARylated nucleosome to the tight binding and activation of the remodeler. Functional biochemical assays with PARylated nucleosomes highlight the importance of nucleosomal epitopes for productive remodeling and suggest that ALC1 preferentially slides nucleosomes away from DNA breaks.",
author = "Luka Bacic and Guillaume Gaullier and Anton Sabantsev and Lehmann, {Laura C.} and Klaus Brackmann and Despoina Dimakou and Mario Halic and Graeme Hewitt and Boulton, {Simon J.} and Sebastian Deindl",
note = "Funding Information: Cryo-EM data were collected at the Swedish National Cryo-EM Facility funded by the Knut and Alice Wallenberg, Family Erling Persson and Kempe Foundations, SciLifeLab, Stockholm University, and Ume{\aa} University. We thank M Carroni, K Wallden, and J Conrad for assistance with cryo-EM data collection, and H Sch{\"u}ler for discussions and for providing reagents. We thank J Pascal for generously providing the PARP1 expression vector. The HPF1 expression vector was a kind gift from K Luger. We thank M Schattenhofer for assistance with figure layout. This work was supported by the European Research Council (ERC Starting Grant 714068), the Knut and Alice Wallenberg Foundation (Grant KAW 019.0306), the Swedish Research Council (VR Grant 2019–03534), and Cancerfonden (Grant 19 0055 Pj). SD is an EMBO Young Investigator. GH and SJB are 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); SJB is also funded by a European Research Council (ERC) Advanced Investigator Grant (TelMetab) and Wellcome Trust Senior Investigator and Collaborative Grants. Funding Information: Cryo-EM data were collected at the Swedish National Cryo-EM Facility funded by the Knut and Alice Wallenberg, Family Erling Persson and Kempe Foundations, SciLifeLab, Stockholm University, and Ume? University. We thank M Carroni, K Wallden, and J Conrad for assistance with cryo-EM data collection, and H Sch?ler for discussions and for providing reagents. We thank J Pascal for generously providing the PARP1 expression vector. The HPF1 expression vector was a kind gift from K Luger. We thank M Schattenhofer for assistance with figure layout. This work was supported by the European Research Council (ERC Starting Grant 714068), the Knut and Alice Wallenberg Foundation (Grant KAW 019.0306), the Swedish Research Council (VR Grant 2019?03534), and Cancerfonden (Grant 19 0055 Pj). SD is an EMBO Young Investigator. GH and SJB are 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); SJB is also funded by a European Research Council (ERC) Advanced Investigator Grant (TelMetab) and Wellcome Trust Senior Investigator and Collaborative Grants. Publisher Copyright: {\textcopyright} Bacic et al.",
year = "2021",
month = sep,
doi = "10.7554/eLife.71420",
language = "English",
volume = "10",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications, Ltd",
}