A FRET biosensor for necroptosis uncovers two different modes of the release of DAMPs
AuthorMurai, S; Yamaguchi, Y; Shirasaki, Y; Yamagishi, M; Shindo, R; Hildebrand, JM; Miura, R; Nakabayashi, O; Totsuka, M; Tomida, T; ...
Source TitleNature Communications
PublisherNATURE PUBLISHING GROUP
AffiliationMedical Biology (W.E.H.I.)
Document TypeJournal Article
CitationsMurai, S., Yamaguchi, Y., Shirasaki, Y., Yamagishi, M., Shindo, R., Hildebrand, J. M., Miura, R., Nakabayashi, O., Totsuka, M., Tomida, T., Adachi-Akahane, S., Uemura, S., Silke, J., Yagita, H., Miura, M. & Nakano, H. (2018). A FRET biosensor for necroptosis uncovers two different modes of the release of DAMPs. NATURE COMMUNICATIONS, 9 (1), https://doi.org/10.1038/s41467-018-06985-6.
Access StatusOpen Access
Necroptosis is a regulated form of necrosis that depends on receptor-interacting protein kinase (RIPK)3 and mixed lineage kinase domain-like (MLKL). While danger-associated molecular pattern (DAMP)s are involved in various pathological conditions and released from dead cells, the underlying mechanisms are not fully understood. Here we develop a fluorescence resonance energy transfer (FRET) biosensor, termed SMART (a sensor for MLKL activation by RIPK3 based on FRET). SMART is composed of a fragment of MLKL and monitors necroptosis, but not apoptosis or necrosis. Mechanistically, SMART monitors plasma membrane translocation of oligomerized MLKL, which is induced by RIPK3 or mutational activation. SMART in combination with imaging of the release of nuclear DAMPs and Live-Cell Imaging for Secretion activity (LCI-S) reveals two different modes of the release of High Mobility Group Box 1 from necroptotic cells. Thus, SMART and LCI-S uncover novel regulation of the release of DAMPs during necroptosis.
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