Co-operative and Hierarchical Binding of c-FLIP and Caspase-8: A Unified Model Defines How c-FLIP Isoforms Differentially Control Cell Fate
Open accessAbstract
The death-inducing signaling complex (DISC) initiates death receptor-induced apoptosis. DISC assembly and activation are controlled by c-FLIP isoforms, which function as pro-apoptotic (c-FLIPL only) or anti-apoptotic (c-FLIPL/c-FLIPS) regulators of procaspase-8 activation. Current models assume that c-FLIP directly competes with procaspase-8 for recruitment to FADD. Using a functional reconstituted DISC, structure-guided mutagenesis, and quantitative LC-MS/MS, we show that c-FLIPL/S binding to the DISC is instead a co-operative procaspase-8-dependent process. FADD initially recruits procaspase-8, which in turn recruits and heterodimerizes with c-FLIPL/S via a hierarchical binding mechanism. Procaspase-8 activation is regulated by the ratio of unbound c-FLIPL/S to procaspase-8, which determines composition of the procaspase-8:c-FLIPL/S heterodimer. Thus, procaspase-8:c-FLIPL exhibits localized enzymatic activity and is preferentially an activator, promoting DED-mediated procaspase-8 oligomer assembly, whereas procaspase-8:c-FLIPS lacks activity and potently blocks procaspase-8 activation. This co-operative hierarchical binding model explains the dual role of c-FLIPL and crucially defines how c-FLIP isoforms differentially control cell fate.
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.
Integrity signals
Registry
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.
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?
Heterodimeric complexes of CASP8 with c-FLIPL (but not c-FLIPS) have limited enzymatic activity that favors CASP8 oligomerization and activation.
“heterodimeric complexes of CASP8 with c-FLIPL (but not c-FLIPS) are endowed with limited enzymatic activity that favors CASP8 oligomerization and consequent activation [301].”
From Molecular mechanisms of cell death: recommendations of the nomenclature committee on cell death 2018. (2018) · cites this paper as [301]
c-FLIPS inhibits and c-FLIPL activates CASP8 by modulating CASP8 oligomerization.
“Compelling evidence indicates that the short variant of c-FLIP (c-FLIPS) and its long counterpart (c-FLIPL) inhibit and activate CASP8, respectively, by modulating CASP8 oligomerization [298–301].”
From Molecular mechanisms of cell death: recommendations of the nomenclature committee on cell death 2018. (2018) · cites this paper as [301]
Elevated expression levels of c-FLIPL inhibit, rather than activate, extrinsic apoptosis possibly by disrupting CASP8 maturation.
“supporting the notion that elevated expression levels of c-FLIPL inhibit, rather than activate, extrinsic apoptosis possibly by disrupting CASP8 maturation [301, 304].”
From Molecular mechanisms of cell death: recommendations of the nomenclature committee on cell death 2018. (2018) · cites this paper as [301]
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
Inhibition of death receptor signals by cellular FLIP
Irmler, M., Thome, M., Hahne, M., Schneider, P., Hofmann, K., Steiner, V., Bodmer, J.-L., Schröter, M., Burns, K., Mattmann, C., et al. (1997) · Nature · doi:10.1038/40657 · Cited by 2,548 · see citing works
Kischkel, F. C., Hellbardt, S., Behrmann, I., Germer, M., Pawlita, M., Krammer, P. H., & Peter, M. E. (1995) · The EMBO Journal · doi:10.1002/j.1460-2075.1995.tb00245.x · Cited by 2,181 · see citing works
Absolute Quantification of Proteins by LCMSE
Silva, J. C., Gorenstein, M. V., Li, G.-Z., Vissers, J. P. C., & Geromanos, S. J. (2006) · Molecular & Cellular Proteomics · doi:10.1074/mcp.m500230-mcp200 · Cited by 1,529 · see citing works
Varfolomeev, E., Schuchmann, M., Luria, V., Chiannilkulchai, N., Beckmann, J., Mett, I., Rebrikov, D., Brodianski, V. M., Kemper, O., Kollet, O., et al. (1998) · Immunity · doi:10.1016/s1074-7613(00)80609-3 · Cited by 1,332 · see citing works
Catalytic activity of the caspase-8–FLIPL complex inhibits RIPK3-dependent necrosis
Oberst, A., Dillon, C. P., Weinlich, R., McCormick, L. L., Fitzgerald, P., Pop, C., Hakem, R., Salvesen, G. S., & Green, D. R. (2011) · Nature · doi:10.1038/nature09852 · Cited by 1,278 · cited 4× · see citing works
“Hence, during both development and immune cell proliferation, c-FLIPL:procaspase-8 heterodimers function to inhibit RIPK1RIPK3-mediated programmed necrosis (Oberst et al., 2011).”
InfluentialRIP3 mediates the embryonic lethality of caspase-8-deficient mice
Kaiser, W. J., Upton, J. W., Long, A. B., Livingston-Rosanoff, D., Daley-Bauer, L. P., Hakem, R., Caspary, T., & Mocarski, E. S. (2011) · Nature · doi:10.1038/nature09857 · Cited by 1,057 · see citing works
Feoktistova, M., Geserick, P., Kellert, B., Panayotova‐Dimitrova, D., Langlais, C., Hupe, M., Cain, K., MacFarlane, M., Häcker, G. A., & Leverkus, M. (2011) · Molecular Cell · doi:10.1016/j.molcel.2011.06.011 · Cited by 855 · see citing works
Self-citationThe Role of c-FLIP in Modulation of CD95-induced Apoptosis
Scaffidi, C., Schmitz, I., Krammer, P. H., & Peter, M. E. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.3.1541 · Cited by 819 · see citing works
Tenev, T., Bianchi, K., Darding, M., Broemer, M., Langlais, C., Wållberg, F., Zachariou, A., Lopez, J. S., MacFarlane, M., Cain, K., et al. (2011) · Molecular Cell · doi:10.1016/j.molcel.2011.06.006 · Cited by 818 · see citing works
Self-citationKrueger, A., Schmitz, I., Baumann, S., Krammer, P. H., & Kirchhoff, S. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m101780200 · Cited by 565 · see citing works
c‐FLIPL is a dual function regulator for caspase‐8 activation and CD95‐mediated apoptosis
Chang, D. W.-W., Xing, Z., Pan, Y., Algeciras‐Schimnich, A., Barnhart, B. C., Yaish‐Ohad, S., Peter, M. E., & Yang, X. (2002) · The EMBO Journal · doi:10.1093/emboj/cdf356 · Cited by 560 · see citing works
FLICE-Inhibitory Proteins: Regulators of Death Receptor-Mediated Apoptosis
Krueger, A., Baumann, S., Krammer, P. H., & Kirchhoff, S. (2001) · Molecular and Cellular Biology · doi:10.1128/mcb.21.24.8247-8254.2001 · Cited by 548 · see citing works
Essential role for caspase 8 in T-cell homeostasis and T-cell-mediated immunity.Preprint
Salmena, L., Lemmers, B., Hakem, A., Matysiak-Zablocki, E., Murakami, K., Au, P. Y. B., Berry, D., Tamblyn, L., Shehabeldin, A., Migon, E., et al. (2003) · Genes & Development · doi:10.1101/gad.1063703 · Cited by 517 · see citing works
The Long Form of FLIP Is an Activator of Caspase-8 at the Fas Death-inducing Signaling Complex
Micheau, O., Thome, M., Schneider, P., Holler, N., Tschopp, J., Nicholson, D. W., Briand, C., & Grütter, M. G. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m206882200 · Cited by 482 · see citing works
Many stimuli pull the necrotic trigger, an overview
Vanlangenakker, N., Berghe, T. V., & Vandenabeele, P. (2011) · Cell Death and Differentiation · doi:10.1038/cdd.2011.164 · Cited by 407 · see citing works
Casper Is a FADD- and Caspase-Related Inducer of Apoptosis
Shu, H., Halpin, D. R., & Goeddel, D. V. (1997) · Immunity · doi:10.1016/s1074-7613(00)80450-1 · Cited by 399 · see citing works
The Fas–FADD death domain complex structure unravels signalling by receptor clustering
Scott, F. L., Stec, B., Pop, C., Dobaczewska, M. K., Lee, J. J., Monosov, E., Robinson, H., Salvesen, G. S., Schwarzenbacher, R., & Riedl, S. J. (2009) · Nature · doi:10.1038/nature07606 · Cited by 384 · see citing works
Regulation of lymphocyte proliferation and death by flip
Thome, M., & Tschopp, J. (2001) · Nature reviews. Immunology · doi:10.1038/35095508 · Cited by 375 · see citing works
Dickens, L. S., Boyd, R. S., Jukes-Jones, R., Hughes, M. A., Robinson, G. L., Fairall, L., Schwabe, J. W. R., Cain, K., & MacFarlane, M. (2012) · Molecular Cell · doi:10.1016/j.molcel.2012.05.004 · Cited by 349 · see citing works
Self-citationSurvival Function of the FADD-CASPASE-8-cFLIPL Complex
Dillon, C. P., Oberst, A., Weinlich, R., Janke, L. J., Kang, T.-B., Ben-Moshe, T., Mak, T. W., Wallach, D., & Green, D. R. (2012) · Cell Reports · doi:10.1016/j.celrep.2012.03.010 · Cited by 331 · cited 3× · see citing works
“In addition to its key role in apoptosis, caspase-8 has a survival role as it is required for embryonic development (Dillon et al., 2012; Varfolomeev et al., 1998), immune cell proliferation (Salmena et al.”
Rasper, D., Vaillancourt, J. P., Hadano, S., Houtzager, V. M., Seiden, I. M., Keen, S., Tawa, P., Xanthoudakis, S., Nasir, J., Martindale, D. W., et al. (1998) · Cell Death and Differentiation · doi:10.1038/sj.cdd.4400370 · Cited by 316 · see citing works
The Fas–FADD death domain complex structure reveals the basis of DISC assembly and disease mutations
Wang, L., Yang, J. K., Kabaleeswaran, V., Rice, A. J., Cruz, A. C., Park, A. Y., Yin, Q., Damko, E., Jang, S. B., Raunser, S., et al. (2010) · Nature Structural & Molecular Biology · doi:10.1038/nsmb.1920 · Cited by 276 · see citing works
Activation of caspases-8 and -10 by FLIPL
Boatright, K. M., Cristina, D. E. I. S., Denault, J., Sutherlin, D. P., & Salvesen, G. S. (2004) · Biochemical Journal · doi:10.1042/bj20040809 · Cited by 242 · see citing works
Death-effector Filaments: Novel Cytoplasmic Structures that Recruit Caspases and Trigger Apoptosis
Siegel, R. M., Martin, D. A., Zheng, L., Ng, S. Y., Bertin, J., Cohen, J. I., & Lenardo, M. J. (1998) · The Journal of Cell Biology · doi:10.1083/jcb.141.5.1243 · Cited by 235 · see citing works
Pop, C., Oberst, A., Drąg, M., van Raam, B. J., Riedl, S. J., Green, D. R., & Salvesen, G. S. (2011) · Biochemical Journal · doi:10.1042/bj20101738 · Cited by 225 · see citing works
Oberst, A., Pop, C., Tremblay, A. G., Blais, V., Denault, J.-B., Salvesen, G. S., & Green, D. R. (2010) · Journal of Biological Chemistry · doi:10.1074/jbc.m109.095083 · Cited by 214 · see citing works
Harper, N. W., Hughes, M. A., MacFarlane, M., & Cohen, G. M. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m303399200 · Cited by 209 · see citing works
Self-citationThe ‘complexities’ of life and death: Death receptor signalling platforms
Dickens, L. S., Powley, I. R., Hughes, M. A., & MacFarlane, M. (2012) · Experimental Cell Research · doi:10.1016/j.yexcr.2012.04.005 · Cited by 208 · see citing works
Self-citationSchleich, K., Warnken, U., Fricker, N., Öztürk, S., Richter, P., Kammerer, K., Schnölzer, M., Krammer, P. H., & Lavrik, I. N. (2012) · Molecular Cell · doi:10.1016/j.molcel.2012.05.006 · Cited by 198 · see citing works
Hughes, M. A., Harper, N. W., Butterworth, M., Cain, K., Cohen, G. M., & MacFarlane, M. (2009) · Molecular Cell · doi:10.1016/j.molcel.2009.06.012 · Cited by 185 · see citing works
Self-citationMechanism of procaspase-8 activation by c-FLIP L
Yu, J. W., Jeffrey, P. D., & Shi, Y. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0812453106 · Cited by 157 · see citing works
MacFarlane, M., Merrison, W., Dinsdale, D. D., & Cohen, G. M. (2000) · The Journal of Cell Biology · doi:10.1083/jcb.148.6.1239 · Cited by 157 · see citing works
Self-citationFricker, N., Beaudouin, J., Richter, P., Eils, R., Krammer, P. H., & Lavrik, I. N. (2010) · Journal of Cell Biology · doi:10.1083/jcb.201002060 · Cited by 154 · see citing works
Crystal Structure of MC159 Reveals Molecular Mechanism of DISC Assembly and FLIP Inhibition
Yang, J. K., Wang, L., Zheng, L., Wan, F., Ahmed, M., Lenardo, M. J., & Wu, H. (2005) · Molecular Cell · doi:10.1016/j.molcel.2005.10.023 · Cited by 140 · see citing works
Cellular FLICE-like inhibitory proteins (c-FLIPs): Fine-tuners of life and death decisions
Öztürk, S., Schleich, K., & Lavrik, I. N. (2012) · Experimental Cell Research · doi:10.1016/j.yexcr.2012.01.019 · Cited by 112 · see citing works
Kavuri, S. M., Geserick, P., Berg, D., Dimitrova, D. P., Feoktistova, M., Siegmund, D., Gollnick, H., Neumann, M., Wajant, H., & Leverkus, M. (2011) · Journal of Biological Chemistry · doi:10.1074/jbc.m110.148585 · Cited by 92 · see citing works
Self-citationThe Inflammasome Adaptor ASC Induces Procaspase-8 Death Effector Domain Filaments
Vajjhala, P. R., Lu, A. Z., Brown, D. L., Pang, S. W., Sagulenko, V., Sester, D. P., Cridland, S. O., Hill, J. M., Schroder, K., Stow, J. L., et al. (2015) · Journal of Biological Chemistry · doi:10.1074/jbc.m115.687731 · Cited by 92 · see citing works
Structural and Biochemical Studies on Procaspase-8: New Insights on Initiator Caspase Activation
Keller, N., Mareš, J., Zerbe, O., & Grütter, M. G. (2009) · Structure · doi:10.1016/j.str.2008.12.019 · Cited by 89 · see citing works
Majkut, J., Sgobba, M., Holohan, C., Crawford, N., Logan, A. E., Kerr, E., Higgins, C. A., Redmond, K. L., Riley, J. S., Stasik, I., et al. (2014) · Nature Communications · doi:10.1038/ncomms4350 · Cited by 81 · see citing works
Pick your poison: The Ripoptosome, a cell death platform regulating apoptosis and necroptosis
Feoktistova, M., Geserick, P., Panayotova‐Dimitrova, D., & Leverkus, M. (2012) · Cell Cycle · doi:10.4161/cc.11.3.19060 · Cited by 70 · see citing works
Self-citationEsposito, D., Sankar, A., Morgner, N., Robinson, C. V., Rittinger, K., & Driscoll, P. C. (2010) · Structure · doi:10.1016/j.str.2010.08.006 · Cited by 57 · see citing works
Koenig, A., Buskiewicz, I. A., Fortner, K. A., Russell, J. Q., Asaoka, T., He, Y.-W., Hakem, R., Eriksson, J. E., & Budd, R. C. (2014) · Journal of Biological Chemistry · doi:10.1074/jbc.m113.506428 · Cited by 42 · see citing works
Crystal structure of the death effector domains of caspase-8
Shen, C., Yue, H., Pei, J., Guo, X., Wang, T., & Quan, J. (2015) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2015.05.054 · Cited by 25 · see citing works
c‐FLIPS reduces activation of caspase and NF‐κB pathways and decreases T cell survival
Hinshaw-Makepeace, J. A., Huston, G. E., Fortner, K. A., Russell, J. Q., Holoch, D., Swain, S. L., & Budd, R. C. (2007) · European Journal of Immunology · doi:10.1002/eji.200636956 · Cited by 20 · see citing works
Isolation, characterisation and reconstitution of cell death signalling complexes
Hughes, M. A., Langlais, C., Cain, K., et al. (2013) · Methods · doi:10.1016/j.ymeth.2013.02.006 · Cited by 19 · see citing works
Self-citation
Flagged first, then by citations
Cited by
Sorted by citations
Galluzzi, L., Vitale, I., Aaronson, S. A., Abrams, J. M., Adam, D., Agostinis, P., Alnemri, E. S., Altucci, L., Amelio, I., Andrews, D. W., et al. (2018) · Cell Death & Differentiation · doi:10.1038/s41418-017-0012-4 · Cited by 6,680 · see citing works
Self-citationFu, T.-M., Li, Y., Lu, A., Li, Z., Vajjhala, P. R., Cruz, A. C., Srivastava, D. B., DiMaio, F., Penczek, P. A., Siegel, R. M., et al. (2016) · Molecular Cell · doi:10.1016/j.molcel.2016.09.009 · Cited by 196 · see citing works
Horn, S., Hughes, M. A., Schilling, R., Sticht, C., Tenev, T., Ploesser, M., Meier, P., Sprick, M. R., MacFarlane, M., & Leverkus, M. (2017) · Cell Reports · doi:10.1016/j.celrep.2017.04.010 · Cited by 119 · see citing works
Self-citationThe linear ubiquitin chain assembly complex regulates TRAIL‐induced gene activation and cell death
Lafont, E., Kantari‐Mimoun, C., Draber, P., De Miguel, D., Hartwig, T., Reichert, M., Kupka, S., Shimizu, Y., Taraborrelli, L., Spit, M., et al. (2017) · The EMBO Journal · doi:10.15252/embj.201695699 · Cited by 114 · see citing works
Dual role of DR5 in death and survival signaling leads to TRAIL resistance in cancer cells
Shlyakhtina, Y., Pavet, V., & Gronemeyer, H. (2017) · Cell Death & Disease · doi:10.1038/cddis.2017.423 · Cited by 56 · see citing works
- Authors
- Hughes, M. A., Powley, I. R., Jukes-Jones, R., Horn, S., Feoktistova, M., Fairall, L., Schwabe, J. W. R., Leverkus, M., Cain, K., & MacFarlane, M.
- Affiliations
- MRC Toxicology Unit (GB)
- Heidelberg University (DE)
- RWTH Aachen University (DE)
- University of Leicester (GB)
- Corresponding author
- Cain, Kelvin
- MacFarlane, Marion
- ORCID
- Venue
- Molecular Cell
- Publisher
- Elsevier BV
- Published
- 2016-03
- Volume
- 61
- Issue
- 6
- Pages
- 834-849
- ISSN
- 1097-2765 (ISSN-L)
- PMC
- PMC4819448
- OpenAlex
- W2300529787
- Cited by
- 257
- References
- 45
- Citations by year
- 2026: 8 · 2025: 16 · 2024: 27 · 2023: 20 · 2022: 27 · 2021: 34 · 2020: 40 · 2019: 32 · 2018: 18 · 2017: 25 · 2016: 9 · 2014: 1
- FWCI
- 14.55
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- Molecular Cell (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Figshare (submittedVersion, cc-by)
- Leicester Research Archive (University of Leicester) (submittedVersion, cc-by)
- RWTH Publications (RWTH Aachen) (submittedVersion, other-oa)
- Funders
- Deutsche Forschungsgemeinschaft (RTG2099)
- Medical Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Keywords
- c-FLIPL, c-FLIP, caspase-8 activation, death-inducing signaling complex, FADD, cell fate regulation
- MeSH terms
- Humans, Protein Binding, Mutagenesis, Apoptosis, Cell Lineage, Protein Isoforms, Caspase 8, Fas-Associated Death Domain Protein, CASP8 and FADD-Like Apoptosis Regulating Protein, Tandem Mass Spectrometry
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
