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Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis

Programmed cell death (PCD) is used by plants for development and survival to biotic and abiotic stresses. The role of caspases in PCD is well established in animal cells. Over the past 15 years, the importance of caspase-3-like enzymatic activity for plant PCD completion has been widely documented...

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Autores principales: Ge, Y, Cai, Y-M, Bonneau, L, Rotari, V, Danon, A, McKenzie, E A, McLellan, H, Mach, L, Gallois, P
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5072426/
https://www.ncbi.nlm.nih.gov/pubmed/27058316
http://dx.doi.org/10.1038/cdd.2016.34
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author Ge, Y
Cai, Y-M
Bonneau, L
Rotari, V
Danon, A
McKenzie, E A
McLellan, H
Mach, L
Gallois, P
author_facet Ge, Y
Cai, Y-M
Bonneau, L
Rotari, V
Danon, A
McKenzie, E A
McLellan, H
Mach, L
Gallois, P
author_sort Ge, Y
collection PubMed
description Programmed cell death (PCD) is used by plants for development and survival to biotic and abiotic stresses. The role of caspases in PCD is well established in animal cells. Over the past 15 years, the importance of caspase-3-like enzymatic activity for plant PCD completion has been widely documented despite the absence of caspase orthologues. In particular, caspase-3 inhibitors blocked nearly all plant PCD tested. Here, we affinity-purified a plant caspase-3-like activity using a biotin-labelled caspase-3 inhibitor and identified Arabidopsis thaliana cathepsin B3 (AtCathB3) by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Consistent with this, recombinant AtCathB3 was found to have caspase-3-like activity and to be inhibited by caspase-3 inhibitors. AtCathepsin B triple-mutant lines showed reduced caspase-3-like enzymatic activity and reduced labelling with activity-based caspase-3 probes. Importantly, AtCathepsin B triple mutants showed a strong reduction in the PCD induced by ultraviolet (UV), oxidative stress (H(2)O(2), methyl viologen) or endoplasmic reticulum stress. Our observations contribute to explain why caspase-3 inhibitors inhibit plant PCD and provide new tools to further plant PCD research. The fact that cathepsin B does regulate PCD in both animal and plant cells suggests that this protease may be part of an ancestral PCD pathway pre-existing the plant/animal divergence that needs further characterisation.
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spelling pubmed-50724262016-10-31 Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis Ge, Y Cai, Y-M Bonneau, L Rotari, V Danon, A McKenzie, E A McLellan, H Mach, L Gallois, P Cell Death Differ Original Paper Programmed cell death (PCD) is used by plants for development and survival to biotic and abiotic stresses. The role of caspases in PCD is well established in animal cells. Over the past 15 years, the importance of caspase-3-like enzymatic activity for plant PCD completion has been widely documented despite the absence of caspase orthologues. In particular, caspase-3 inhibitors blocked nearly all plant PCD tested. Here, we affinity-purified a plant caspase-3-like activity using a biotin-labelled caspase-3 inhibitor and identified Arabidopsis thaliana cathepsin B3 (AtCathB3) by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Consistent with this, recombinant AtCathB3 was found to have caspase-3-like activity and to be inhibited by caspase-3 inhibitors. AtCathepsin B triple-mutant lines showed reduced caspase-3-like enzymatic activity and reduced labelling with activity-based caspase-3 probes. Importantly, AtCathepsin B triple mutants showed a strong reduction in the PCD induced by ultraviolet (UV), oxidative stress (H(2)O(2), methyl viologen) or endoplasmic reticulum stress. Our observations contribute to explain why caspase-3 inhibitors inhibit plant PCD and provide new tools to further plant PCD research. The fact that cathepsin B does regulate PCD in both animal and plant cells suggests that this protease may be part of an ancestral PCD pathway pre-existing the plant/animal divergence that needs further characterisation. Nature Publishing Group 2016-09-01 2016-04-08 /pmc/articles/PMC5072426/ /pubmed/27058316 http://dx.doi.org/10.1038/cdd.2016.34 Text en Copyright © 2016 Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Original Paper
Ge, Y
Cai, Y-M
Bonneau, L
Rotari, V
Danon, A
McKenzie, E A
McLellan, H
Mach, L
Gallois, P
Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis
title Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis
title_full Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis
title_fullStr Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis
title_full_unstemmed Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis
title_short Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis
title_sort inhibition of cathepsin b by caspase-3 inhibitors blocks programmed cell death in arabidopsis
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5072426/
https://www.ncbi.nlm.nih.gov/pubmed/27058316
http://dx.doi.org/10.1038/cdd.2016.34
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