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The adaptor protein CRK is a pro-apoptotic transducer of endoplasmic reticulum stress

Nature Cell Biology volume 14, pages 87–92 (2012)Cite this article

Abstract

Excessive demands on the protein-folding capacity of the endoplasmic reticulum (ER) cause irremediable ER stress and contribute to cell loss in a number of cell degenerative diseases, including type 2 diabetes and neurodegeneration1,2. The signals communicating catastrophic ER damage to the mitochondrial apoptotic machinery remain poorly understood3,4,5,6. We used a biochemical approach to purify a cytosolic activity induced by ER stress that causes release of cytochrome c from isolated mitochondria. We discovered that the principal component of the purified pro-apoptotic activity is the proto-oncoprotein CRK (CT10-regulated kinase), an adaptor protein with no known catalytic activity7. C r k−/− cells are strongly resistant to ER-stress-induced apoptosis. Moreover, CRK is cleaved in response to ER stress to generate an amino-terminal Mr∼14K fragment with greatly enhanced cytotoxic potential. We identified a putative BH3 (BCL2 homology 3) domain within this N-terminal CRK fragment, which sensitizes isolated mitochondria to cytochrome c release and when mutated significantly reduces the apoptotic activity of CRK in vivo. Together these results identify CRK as a pro-apoptotic protein that signals irremediable ER stress to the mitochondrial execution machinery.

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Figure 1: Biochemical purification of ER stress apoptotic activity identifies CRK.
Figure 2: C r k−/− MEFs are significantly resistant to ER-stress-induced apoptosis.
Figure 3: CRKI or CRKII restores sensitivity of C r k−/− MEFs to ER-stress-induced apoptosis.
Figure 4: CRK is proteolytically cleaved into an apoptotic signal following irremediable ER stress.
Figure 5: CRKII contains a putative BH3 domain and triggers BAX/BAK-dependent apoptosis.

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Acknowledgements

We thank D. Ganem, F. Papa and W. Greene for scientific advice and encouragement throughout this project. We thank C. Lin for help in preparing figures for the manuscript. We thank C. Crane for quantitative PCR assistance. We thank D. Winant at the Stanford PAN facility and N. Krogan and G. Cagney, UCSF, for mass spectrometry analysis of purified samples. This work was supported by NIH grants K08 AI054650 and RO1 CA136577 (S.A.O.); an HHMI Physician-Scientist Early Career Award (S.A.O.); the Steward Trust Foundation (S.A.O.); and the Sandler Program in Basic Sciences (S.A.O.). This work was also supported by a Pennsylvania Department of Health Cure Formulary grant (SAP#4100047628 to T.C.).

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  1. Department of Pathology, University of California-San Francisco, 513 Parnassus Avenue, HSW-517, Box 0511, San Francisco, California 94143-0511, USA

    Kathryn Austgen, Emily T. Johnson & Scott A. Oakes

  2. Department of Pathology and Laboratory Medicine, The Children’s Hospital of Philadelphia Research Institute, Philadelphia, Pennsylvania 19104, USA

    Tae-Ju Park & Tom Curran

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K.A. and E.T.J. designed and carried out experiments and contributed to the manuscript. T-J.P. and T.C. contributed reagents and data interpretation. S.A.O. designed the study and wrote the manuscript.

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Correspondence to Scott A. Oakes.

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Austgen, K., Johnson, E., Park, TJ. et al. The adaptor protein CRK is a pro-apoptotic transducer of endoplasmic reticulum stress. Nat Cell Biol 14, 87–92 (2012). https://doi.org/10.1038/ncb2395

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