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TitleStructure and dynamics of the chromatin remodeler ALC1 bound to a PARylated nucleosome.
Journal, issue, pagesElife, Vol. 10, Year 2021
Publish dateSep 6, 2021
AuthorsLuka Bacic / Guillaume Gaullier / Anton Sabantsev / Laura C Lehmann / Klaus Brackmann / Despoina Dimakou / Mario Halic / Graeme Hewitt / Simon J Boulton / Sebastian Deindl /
PubMed AbstractThe 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 ...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.
External linksElife / PubMed:34486521 / PubMed Central
MethodsEM (single particle)
Resolution4.1 - 10.0 Å
Structure data

13065

7otq

EMDB-13065, PDB-7otq:
Cryo-EM structure of ALC1/CHD1L bound to a PARylated nucleosome
Method: EM (single particle) / Resolution: 4.8 Å

EMDB-13070:
cryoDRGN graph traversal of the ALC1 - PARylated nucleosome complex particle distribution
Method: EM (single particle) / Resolution: 10.0 Å

EMDB-17944: Cryo-EM structure of an asymmetric nucleosome (10-N-0) from a Glacios/Falcon3EC
Method: EM (single particle) / Resolution: 4.1 Å

Source
  • xenopus laevis (African clawed frog)
  • homo sapiens (human)
  • synthetic construct (others)
KeywordsDNA BINDING PROTEIN / ALC1 / CHD1L / chromatin remodeler / DNA damage response / nucleosome / poly(ADP-ribose)

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