TY - JOUR
T1 - Linear ubiquitination by LUBEL has a role in Drosophila heat stress response
AU - Asaoka, Tomoko
AU - Almagro, Jorge
AU - Ehrhardt, Christine
AU - Tsai, Isabella
AU - Schleiffer, Alexander
AU - Deszcz, Luiza
AU - Junttila, Sini
AU - Ringrose, Leonie
AU - Mechtler, Karl
AU - Kavirayani, Anoop
AU - Gyenesei, Attila
AU - Hofmann, Kay
AU - Duchek, Peter
AU - Rittinger, Katrin
AU - Ikeda, Fumiyo
N1 - Funding Information:
We acknowledge Conchi Martinez and Ryoko Shiraishi (IMBA, Vienna, Austria), Joseph Gokcezade (IMBA Fly house), Richard Imre (Protein chemistry, IMP-IMBA core facilities), Tamara Engelmaier and Agnieszka Piszczek (HistoPathology, The Vienna Biocenter Core Facilities GmbH (VBCF), Vienna, Austria), Peggy Stolt-Bergner, Katharina Radakovics, and Jana Neuhold (Protein Technologies, VBCF) for their technical supports. We also thank Molecular Biology Service and Biooptics (IMP-IMBA core facilities) and Next-Generation Sequencing Core Facility (VBCF) for the sample processing and analysis, Masayuki Miura (Tokyo Univ., Tokyo, Japan) and Francois Bonnay (IMBA, Vienna, Austria) for constructive discussions and advices. We thank all the members in Ikeda Lab for scientific discussions on the project and for comments on the manuscript, and Life Science Editors for editorial assistance. The work was supported by the ERC consolidator grant (LUbi, 614711), OeNB, and COST (European Cooperation in Science and Technology, PROTEOSTASIS BM1307) (F.I.), Austrian Academy of Sciences (F.I. and L.R), the Francis Crick Institute (grant number FCI01) which receives its core funding from Cancer Research UK, the UK Medical Research Council, and the Wellcome Trust (K.R.), SFB670 DFG (K.H.), the Austrian Science Fund (SFB F3402; P2465-B24; and TRP 308-N15), and MEIOsys (222883-2) (K. M.).
Publisher Copyright:
© 2016 The Authors. Published under the terms of the CC BY 4.0 license
PY - 2016/11/1
Y1 - 2016/11/1
N2 - The HOIP ubiquitin E3 ligase generates linear ubiquitin chains by forming a complex with HOIL-1L and SHARPIN in mammals. Here, we provide the first evidence of linear ubiquitination induced by a HOIP orthologue in Drosophila. We identify Drosophila CG11321, which we named Linear Ubiquitin E3 ligase (LUBEL), and find that it catalyzes linear ubiquitination in vitro. We detect endogenous linear ubiquitin chain-derived peptides by mass spectrometry in Drosophila Schneider 2 cells and adult flies. Furthermore, using CRISPR/Cas9 technology, we establish linear ubiquitination-defective flies by mutating residues essential for the catalytic activity of LUBEL. Linear ubiquitination signals accumulate upon heat shock in flies. Interestingly, flies with LUBEL mutations display reduced survival and climbing defects upon heat shock, which is also observed upon specific LUBEL depletion in muscle. Thus, LUBEL is involved in the heat response by controlling linear ubiquitination in flies.
AB - The HOIP ubiquitin E3 ligase generates linear ubiquitin chains by forming a complex with HOIL-1L and SHARPIN in mammals. Here, we provide the first evidence of linear ubiquitination induced by a HOIP orthologue in Drosophila. We identify Drosophila CG11321, which we named Linear Ubiquitin E3 ligase (LUBEL), and find that it catalyzes linear ubiquitination in vitro. We detect endogenous linear ubiquitin chain-derived peptides by mass spectrometry in Drosophila Schneider 2 cells and adult flies. Furthermore, using CRISPR/Cas9 technology, we establish linear ubiquitination-defective flies by mutating residues essential for the catalytic activity of LUBEL. Linear ubiquitination signals accumulate upon heat shock in flies. Interestingly, flies with LUBEL mutations display reduced survival and climbing defects upon heat shock, which is also observed upon specific LUBEL depletion in muscle. Thus, LUBEL is involved in the heat response by controlling linear ubiquitination in flies.
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U2 - 10.15252/embr.201642378
DO - 10.15252/embr.201642378
M3 - Article
C2 - 27702987
AN - SCOPUS:84990228262
SN - 1469-221X
VL - 17
SP - 1624
EP - 1640
JO - EMBO Reports
JF - EMBO Reports
IS - 11
ER -