Condos, T. E.C., Dunkerley, K. M., Freeman, E. A., Barber, K. R., Aguirre, J. D., Chaugule, V. K., Xiao, Y., Konermann, L., Walden, H. and Shaw, G. S. (2018) Synergistic recruitment of UbcH7~Ub and phosphorylated Ubl domain triggers parkin activation. EMBO Journal, e100014. (doi: 10.15252/embj.2018100014) (PMID:30446597)
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Abstract
The E3 ligase parkin ubiquitinates outer mitochondrial membrane proteins during oxidative stress and is linked to early-onset Parkinson’s disease. Parkin is autoinhibited but is activated by the kinase PINK1 that phosphorylates ubiquitin leading to parkin recruitment, and stimulates phosphorylation of parkin’s N-terminal ubiquitin-like (pUbl) domain. How these events alter the structure of parkin to allow recruitment of an E2~Ub conjugate and enhanced ubiquitination is an unresolved question. We present a model of an E2~Ub conjugate bound to the phosphoubiquitin-loaded C-terminus of parkin, derived from NMR chemical shift perturbation experiments. We show the UbcH7~Ub conjugate binds in the open state whereby conjugated ubiquitin binds to the RING1/IBR interface. Further, NMR and mass spectrometry experiments indicate the RING0/RING2 interface is re-modelled, remote from the E2 binding site, and this alters the reactivity of the RING2(Rcat) catalytic cysteine, needed for ubiquitin transfer. Our experiments provide evidence that parkin phosphorylation and E2~Ub recruitment act synergistically to enhance a weak interaction of the pUbl domain with the RING0 domain and rearrange the location of the RING2(Rcat) domain to drive parkin activity.
| Item Type: | Articles |
|---|---|
| Additional Information: | This work was supported by postgraduate scholarships from the Natural Sciences and Engineering Research Council of Canada (TECC, KMD), a collaborative travel grant from Boehringer-Ingelheim Fonds (TECC), research grants from the Canadian Institutes of Health Research (MOP-14606) to GSS, Medical Research Council (MC_UU_12016/12) to HW and Natural Sciences and Engineering Research Council of Canada (RGPIN-2018-04243) to LK, and a Wellcome Trust Investigator Award (209347/Z/17/Z) to HW. |
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Chaugule, Dr Viduth and Walden, Professor Helen |
| Authors: | Condos, T. E.C., Dunkerley, K. M., Freeman, E. A., Barber, K. R., Aguirre, J. D., Chaugule, V. K., Xiao, Y., Konermann, L., Walden, H., and Shaw, G. S. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Molecular Biosciences |
| Journal Name: | EMBO Journal |
| Publisher: | EMBO Press |
| ISSN: | 0261-4189 |
| ISSN (Online): | 1460-2075 |
| Copyright Holders: | Copyright © 2018 The Authors |
| First Published: | First published in EMBO Journal e100014 |
| Publisher Policy: | Reproduced under a Creative Commons License |
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