CoChat
Sign Up
Cited by 406Cell death mechanisms and regulationInflammasome and immune disordersCancer-related Molecular Pathways

Recruitment, activation and retention of caspases‐9 and ‐3 by Apaf‐1 apoptosome and associated XIAP complexes

Open access
Bratton, Shawn B., Walker, Gail, Srinivasula, Srinivasa M., Sun, Xiao‐Ming, Butterworth, Michael, Alnemri, Emad S., Cohen, Gerald M.·2001·The EMBO Journal
DOI 10.1093/emboj/20.5.998PMC PMC145489
Open this paper in CoChat — free account

Abstract

During apoptosis, release of cytochrome c initiates dATP-dependent oligomerization of Apaf-1 and formation of the apoptosome. In a cell-free system, we have addressed the order in which apical and effector caspases, caspases-9 and -3, respectively, are recruited to, activated and retained within the apoptosome. We propose a multi-step process, whereby catalytically active processed or unprocessed caspase-9 initially binds the Apaf-1 apoptosome in cytochrome c/dATP-activated lysates and consequently recruits caspase-3 via an interaction between the active site cysteine (C287) in caspase-9 and a critical aspartate (D175) in caspase-3. We demonstrate that XIAP, an inhibitor-of-apoptosis protein, is normally present in high molecular weight complexes in unactivated cell lysates, but directly interacts with the apoptosome in cytochrome c/dATP-activated lysates. XIAP associates with oligomerized Apaf-1 and/or processed caspase-9 and influences the activation of caspase-3, but also binds activated caspase-3 produced within the apoptosome and sequesters it within the complex. Thus, XIAP may regulate cell death by inhibiting the activation of caspase-3 within the apoptosome and by preventing release of active caspase-3 from the complex.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1093/emboj/20.5.998

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • APAF1 is the molecular core of the apoptosome and is typically required for activation of the caspases that initiate apoptosis.

    “APAF1 (apoptotic protease activating factor 1) is the molecular core of apoptosome. It is typically required for activation of the caspases that initiate apoptosis (3, 26).”

    From MicroRNA 21 Promotes Glioma Invasion by Targeting Matrix Metalloproteinase Regulators (2008) · cites this paper as [3]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

No references could be resolved yet.

Research works printed but not linked: 37

  1. Bratton S.B., MacFarlane,M., Cain,K. and Cohen,G.M. (2000) Protein complexes activate distinct caspase cascades in death receptor and stress-induced apoptosis. Exp. Cell Res., 256, 27–33. [DOI] [PubMed] [Google Scholar]
  2. Cain K., Brown,D.G., Langlais,C. and Cohen,G.M. (1999) Caspase activation involves the formation of the aposome, a large (∼700 kDa) caspase-activating complex. J. Biol. Chem., 274, 22686–22692. [DOI] [PubMed] [Google Scholar]
  3. Cain K., Bratton,S.B., Langlais,C., Walker,G., Brown,D.G., Sun,X.M. and Cohen,G.M. (2000) Apaf-1 oligomerizes into biologically active ∼700 kDa and inactive ∼1.4 MDa apoptosome complexes. J. Biol. Chem., 275, 6067–6070. [DOI] [PubMed] [Google Scholar]
  4. Clem R.J. and Miller,L.K. (1994) Control of programmed cell death by the baculovirus genes p35 and iap. Mol. Cell. Biol., 14, 5212–5222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen G.M. (1997) Caspases: the executioners of apoptosis. Biochem. J., 326, 1–16. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Deveraux Q.L. and Reed,J.C. (1999) IAP family proteins—suppressors of apoptosis. Genes Dev., 13, 239–252. [DOI] [PubMed] [Google Scholar]
  7. Deveraux Q.L., Takahashi,R., Salvesen,G.S. and Reed,J.C. (1997) X-linked IAP is a direct inhibitor of cell-death proteases. Nature, 388, 300–304. [DOI] [PubMed] [Google Scholar]
  8. Deveraux Q.L., Roy,N., Stennicke,H.R., Van Arsdale,T., Zhou,Q., Srinivasula,S.M., Alnemri,E.S., Salvesen,G.S. and Reed,J.C. (1998) IAPs block apoptotic events induced by caspase-8 and cytochrome c by direct inhibition of distinct caspases. EMBO J., 17, 2215–2223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deveraux Q.L., Leo,E., Stennicke,H.R., Welsh,K., Salvesen,G.S. and Reed,J.C. (1999) Cleavage of human inhibitor of apoptosis protein XIAP results in fragments with distinct specificities for caspases. EMBO J., 18, 5242–5251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Earnshaw W.C., Martins,L.M. and Kaufmann,S.H. (1999) Mammalian caspases: structure, activation, substrates and functions during apoptosis. Annu. Rev. Biochem., 68, 383–424. [DOI] [PubMed] [Google Scholar]
  11. Faleiro L. and Lazebnik,Y. (2000) Caspases disrupt the nuclear–cytoplasmic barrier. J. Cell Biol., 151, 951–960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fernandes-Alnemri T. et al. (1996) In vitro activation of CPP32 and Mch3 by Mch4, a novel human apoptotic cysteine protease containing two FADD-like domains. Proc. Natl Acad. Sci. USA, 93, 7464–7469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Green D.R. and Reed,J.C. (1998) Mitochondria and apoptosis. Science, 281, 1309–1312. [DOI] [PubMed] [Google Scholar]
  14. Hakem R. et al. (1998) Differential requirement for caspase 9 in apoptotic pathways in vivo. Cell, 94, 339–352. [DOI] [PubMed] [Google Scholar]
  15. Han Z., Hendrickson,E.A., Bremner,T.A. and Wyche,J.H. (1997) A sequential two-step mechanism for the production of the mature p17:p12 form of caspase-3 in vitro. J. Biol. Chem., 272, 13432–13436. [DOI] [PubMed] [Google Scholar]
  16. Hu Y., Ding,L., Spencer,D.M. and Nunez,G. (1998) WD-40 repeat region regulates Apaf-1 self-association and procaspase-9 activation. J. Biol. Chem., 273, 33489–33494. [DOI] [PubMed] [Google Scholar]
  17. Hu Y., Benedict,M.A., Ding,L. and Nunez,G. (1999) Role of cytochrome c and dATP/ATP hydrolysis in Apaf-1-mediated caspase-9 activation and apoptosis. EMBO J., 18, 3586–3595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kuida K., Haydar,T.F., Kuan,C.Y., Gu,Y., Taya,C., Karasuyama,H., Su,M.S., Rakic,P. and Flavell,R.A. (1998) Reduced apoptosis and cytochrome c-mediated caspase activation in mice lacking caspase 9. Cell, 94, 325–337. [DOI] [PubMed] [Google Scholar]
  19. Li P., Nijhawan,D., Budihardjo,I., Srinivasula,S.M., Ahmad,M., Alnemri,E.S. and Wang,X. (1997) Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell, 91, 479–489. [DOI] [PubMed] [Google Scholar]
  20. Nicholson D.W. et al. (1995) Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature, 376, 37–43. [DOI] [PubMed] [Google Scholar]
  21. Pandey P., Farber,R., Nakazawa,A., Kumar,S., Bharti,A., Nalin,C., Weichselbaum,R., Kufe,D. and Kharbanda,S. (2000) Hsp27 functions as a negative regulator of cytochrome c-dependent activation of procaspase-3. Oncogene, 19, 1975–1981. [DOI] [PubMed] [Google Scholar]
  22. Qin H., Srinivasula,S.M., Wu,G., Fernandes-Alnemri,T., Alnemri,E.S. and Shi,Y. (1999) Structural basis of procaspase-9 recruitment by the apoptotic protease-activating factor 1. Nature, 399, 549–557. [DOI] [PubMed] [Google Scholar]
  23. Rodriguez J. and Lazebnik,Y. (1999) Caspase-9 and APAF-1 form an active holoenzyme. Genes Dev., 13, 3179–3184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Saleh A., Srinivasula,S.M., Acharya,S., Fishel,R. and Alnemri,E.S. (1999) Cytochrome c and dATP-mediated oligomerization of Apaf-1 is a prerequisite for procaspase-9 activation. J. Biol. Chem., 274, 17941–17945. [DOI] [PubMed] [Google Scholar]
  25. Samali A., Cai,J., Zhivotovsky,B., Jones,D.P. and Orrenius,S. (1999) Presence of a pre-apoptotic complex of pro-caspase-3, Hsp60 and Hsp10 in the mitochondrial fraction of Jurkat cells. EMBO J., 18, 2040–2048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Slee E.A. et al. (1999) Ordering the cytochrome c-initiated caspase cascade: hierarchical activation of caspases-2, -3, -6, -7, -8 and -10 in a caspase-9-dependent manner. J. Cell Biol., 144, 281–292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Srinivasula S.M., Ahmad,M., Fernandes-Alnemri,T., Litwack,G. and Alnemri,E.S. (1996) Molecular ordering of the Fas-apoptotic pathway: the Fas/APO-1 protease Mch5 is a CrmA-inhibitable protease that activates multiple Ced-3/ICE-like cysteine proteases. Proc. Natl Acad. Sci. USA, 93, 14486–14491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Srinivasula S.M., Ahmad,M., Fernandes-Alnemri,T. and Alnemri,E.S. (1998) Autoactivation of procaspase-9 by Apaf-1-mediated oligomerization. Mol. Cell, 1, 949–957. [DOI] [PubMed] [Google Scholar]
  29. Stennicke H.R. and Salvesen,G.S. (1999) Catalytic properties of the caspases. Cell Death Differ., 6, 1054–1059. [DOI] [PubMed] [Google Scholar]
  30. Stennicke H.R., Deveraux,Q.L., Humke,E.W., Reed,J.C., Dixit,V.M. and Salvesen,G.S. (1999) Caspase-9 can be activated without proteolytic processing. J. Biol. Chem., 274, 8359–8362. [DOI] [PubMed] [Google Scholar]
  31. Sun C. et al. (1999) NMR structure and mutagenesis of the inhibitor-of-apoptosis protein XIAP. Nature, 401, 818–822. [DOI] [PubMed] [Google Scholar]
  32. Sun C., Cai,M., Meadows,R.P., Xu,N., Gunasekera,A.H., Herrmann,J., Wu,J.C. and Fesik,S.W. (2000) NMR structure and mutagenesis of the third Bir domain of the inhibitor of apoptosis protein XIAP. J. Biol. Chem., 275, 33777–33781. [DOI] [PubMed] [Google Scholar]
  33. Takahashi R., Deveraux,Q., Tamm,I., Welsh,K., Assa-Munt,N., Salvesen,G.S. and Reed,J.C. (1998) A single BIR domain of XIAP sufficient for inhibiting caspases. J. Biol. Chem., 273, 7787–7790. [DOI] [PubMed] [Google Scholar]
  34. Vucic D., Stennicke,H.R., Pisabarro,M.T., Salvesen,G.S. and Dixit,V.M. (2000) ML-IAP, a novel inhibitor of apoptosis that is preferentially expressed in human melanomas. Curr. Biol., 10, 1359–1366. [DOI] [PubMed] [Google Scholar]
  35. Woo M. et al. (1998) Essential contribution of caspase 3/CPP32 to apoptosis and its associated nuclear changes. Genes Dev., 12, 806–819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Zou H., Henzel,W.J., Liu,X., Lutschg,A. and Wang,X. (1997) Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3. Cell, 90, 405–413. [DOI] [PubMed] [Google Scholar]
  37. Zou H., Li,Y., Liu,X. and Wang,X. (1999) An APAF-1⋅cytochrome c multimeric complex is a functional apoptosome that activates procaspase-9. J. Biol. Chem., 274, 11549–11556. [DOI] [PubMed] [Google Scholar]

Cited by

Sorted by citations

Authors
Bratton, S. B., Walker, G., Srinivasula, S. M., Sun, X., Butterworth, M., Alnemri, E. S., & Cohen, G. M.
Affiliations
  • University of Leicester (GB)
  • Thomas Jefferson University (US)
Corresponding author
Cohen, Gerald M.
ORCID
Venue
The EMBO Journal
Publisher
Springer Science and Business Media LLC
Published
2001-03-01
Volume
20
Issue
5
Pages
998-1009
ISSN
  • 0261-4189 (ISSN-L)
  • 1460-2075
DOI
10.1093/emboj/20.5.998
PMC
PMC145489
OpenAlex
W2022569128
Cited by
406
References
0
Citations by year
2026: 4 · 2025: 9 · 2024: 7 · 2023: 7 · 2022: 11 · 2021: 18 · 2020: 19 · 2019: 10 · 2018: 21 · 2017: 14 · 2016: 14 · 2015: 16 · 2014: 9 · 2013: 11 · 2012: 10
FWCI
11.66
Type
journal article
Language
en
Open access
Licence
Locations
  • The EMBO Journal (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Leicester Research Archive (University of Leicester) (submittedVersion)
Funders
  • University of Texas Southwestern Medical Center
  • European Commission (QLG1-1999-00739)
  • National Institutes of Health (5r01ag013847-09)
  • National Institute on Aging (AG-13847)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Inflammasome and immune disorders
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
Keywords
apoptosis, caspase-9, caspase-3, Apaf-1, apoptosome, XIAP, inhibitor of apoptosis proteins, cytochrome c release, caspase activation, cell death regulation
MeSH terms
Aspartic Acid, Cell Line, Deoxyadenine Nucleotides, Enzyme Activation, Humans, Models, Biological, Precipitin Tests, Protein Binding, Protein Processing, Post-Translational, Proteins, Recombinant Proteins, Apoptosis, Caspases, Macromolecular Substances, X-Linked Inhibitor of Apoptosis Protein, Caspase 3, Caspase 9, Apoptotic Protease-Activating Factor 1

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review