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TitleStructure of the apoptosome: mechanistic insights into activation of an initiator caspase from Drosophila.
Journal, issue, pagesGenes Dev, Vol. 29, Issue 3, Page 277-287, Year 2015
Publish dateFeb 1, 2015
AuthorsYuxuan Pang / Xiao-chen Bai / Chuangye Yan / Qi Hao / Zheqin Chen / Jia-Wei Wang / Sjors H W Scheres / Yigong Shi /
PubMed AbstractApoptosis is executed by a cascade of caspase activation. The autocatalytic activation of an initiator caspase, exemplified by caspase-9 in mammals or its ortholog, Dronc, in fruit flies, is ...Apoptosis is executed by a cascade of caspase activation. The autocatalytic activation of an initiator caspase, exemplified by caspase-9 in mammals or its ortholog, Dronc, in fruit flies, is facilitated by a multimeric adaptor complex known as the apoptosome. The underlying mechanism by which caspase-9 or Dronc is activated by the apoptosome remains unknown. Here we report the electron cryomicroscopic (cryo-EM) structure of the intact apoptosome from Drosophila melanogaster at 4.0 Å resolution. Analysis of the Drosophila apoptosome, which comprises 16 molecules of the Dark protein (Apaf-1 ortholog), reveals molecular determinants that support the assembly of the 2.5-MDa complex. In the absence of dATP or ATP, Dronc zymogen potently induces formation of the Dark apoptosome, within which Dronc is efficiently activated. At 4.1 Å resolution, the cryo-EM structure of the Dark apoptosome bound to the caspase recruitment domain (CARD) of Dronc (Dronc-CARD) reveals two stacked rings of Dronc-CARD that are sandwiched between two octameric rings of the Dark protein. The specific interactions between Dronc-CARD and both the CARD and the WD40 repeats of a nearby Dark protomer are indispensable for Dronc activation. These findings reveal important mechanistic insights into the activation of initiator caspase by the apoptosome.
External linksGenes Dev / PubMed:25644603 / PubMed Central
MethodsEM (single particle)
Resolution4.0 - 4.1 Å
Structure data

2870

3j9k

EMDB-2870: Cryo-EM structure of the multimeric complex between Dark and Dronc-CARD.
PDB-3j9k: Structure of Dark apoptosome in complex with Dronc CARD domain
Method: EM (single particle) / Resolution: 4.1 Å

2871

3j9l

EMDB-2871: Cryo-EM structure of the Dark apoptosome.
PDB-3j9l: Structure of Dark apoptosome from Drosophila melanogaster
Method: EM (single particle) / Resolution: 4.0 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

ChemComp-DTP:
2'-DEOXYADENOSINE 5'-TRIPHOSPHATE / Deoxyadenosine triphosphate

Source
  • drosophila melanogaster (fruit fly)
KeywordsAPOPTOSIS / programmed cell death / caspase activation

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