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Dimer-specific immunoprecipitation of active caspase-2 identifies TRAF proteins as novel activators.

Publication ,  Journal Article
Robeson, AC; Lindblom, KR; Wojton, J; Kornbluth, S; Matsuura, K
Published in: The EMBO journal
July 2018

Caspase-2 has been shown to initiate apoptotic cell death in response to specific intracellular stressors such as DNA damage. However, the molecular mechanisms immediately upstream of its activation are still poorly understood. We combined a caspase-2 bimolecular fluorescence complementation (BiFC) system with fluorophore-specific immunoprecipitation to isolate and study the active caspase-2 dimer and its interactome. Using this technique, we found that tumor necrosis factor receptor-associated factor 2 (TRAF2), as well as TRAF1 and 3, directly binds to the active caspase-2 dimer. TRAF2 in particular is necessary for caspase-2 activation in response to apoptotic cell death stimuli. Furthermore, we found that dimerized caspase-2 is ubiquitylated in a TRAF2-dependent manner at K15, K152, and K153, which in turn stabilizes the active caspase-2 dimer complex, promotes its association with an insoluble cellular fraction, and enhances its activity to fully commit the cell to apoptosis. Together, these data indicate that TRAF2 positively regulates caspase-2 activation and consequent cell death by driving its activation through dimer-stabilizing ubiquitylation.

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Published In

The EMBO journal

DOI

EISSN

1460-2075

ISSN

0261-4189

Publication Date

July 2018

Volume

37

Issue

14

Start / End Page

e97072

Related Subject Headings

  • TNF Receptor-Associated Factor 3
  • TNF Receptor-Associated Factor 1
  • Protein Multimerization
  • Protein Interaction Mapping
  • Protein Binding
  • Immunoprecipitation
  • Humans
  • Developmental Biology
  • Cell Line
  • Caspase 2
 

Citation

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Robeson, A. C., Lindblom, K. R., Wojton, J., Kornbluth, S., & Matsuura, K. (2018). Dimer-specific immunoprecipitation of active caspase-2 identifies TRAF proteins as novel activators. The EMBO Journal, 37(14), e97072. https://doi.org/10.15252/embj.201797072
Robeson, Alexander C., Kelly R. Lindblom, Jeffrey Wojton, Sally Kornbluth, and Kenkyo Matsuura. “Dimer-specific immunoprecipitation of active caspase-2 identifies TRAF proteins as novel activators.The EMBO Journal 37, no. 14 (July 2018): e97072. https://doi.org/10.15252/embj.201797072.
Robeson AC, Lindblom KR, Wojton J, Kornbluth S, Matsuura K. Dimer-specific immunoprecipitation of active caspase-2 identifies TRAF proteins as novel activators. The EMBO journal. 2018 Jul;37(14):e97072.
Robeson, Alexander C., et al. “Dimer-specific immunoprecipitation of active caspase-2 identifies TRAF proteins as novel activators.The EMBO Journal, vol. 37, no. 14, July 2018, p. e97072. Epmc, doi:10.15252/embj.201797072.
Robeson AC, Lindblom KR, Wojton J, Kornbluth S, Matsuura K. Dimer-specific immunoprecipitation of active caspase-2 identifies TRAF proteins as novel activators. The EMBO journal. 2018 Jul;37(14):e97072.

Published In

The EMBO journal

DOI

EISSN

1460-2075

ISSN

0261-4189

Publication Date

July 2018

Volume

37

Issue

14

Start / End Page

e97072

Related Subject Headings

  • TNF Receptor-Associated Factor 3
  • TNF Receptor-Associated Factor 1
  • Protein Multimerization
  • Protein Interaction Mapping
  • Protein Binding
  • Immunoprecipitation
  • Humans
  • Developmental Biology
  • Cell Line
  • Caspase 2