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

Y. Ge, Y.-M. Cai, L. Bonneau, V. Rotari, A. Danon, E. A. McKenzie, H. McLellan, L. Mach, P. Gallois (Lead / Corresponding author)

Research output: Contribution to journalArticle

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Abstract

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 (H2O2, 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.

Original languageEnglish
Pages (from-to)1493-1501
Number of pages9
JournalCell Death & Differentiation
Volume23
Issue number9
Early online date8 Apr 2016
DOIs
Publication statusPublished - 1 Sep 2016

Fingerprint

Cathepsin B
Caspase Inhibitors
Arabidopsis
Caspase 3
Cell Death
Plant Cells
Cathepsins
Caspases
Paraquat
Endoplasmic Reticulum Stress
Plant Development
Biotin
Tandem Mass Spectrometry
Liquid Chromatography
Oxidative Stress
Peptide Hydrolases

Keywords

  • Amino acid sequence
  • Apoptosis
  • Arabidopsis
  • Arabidopsis proteins
  • Caspase inhibitors
  • Cathepsin B
  • Chromatography, High Pressure Liquid
  • Endoplasmic reticulum stress
  • Hydrogen peroxide
  • Oxidative stress
  • Paraquat
  • Phylogeny
  • Plants, Genetically modified
  • Recombinant proteins
  • Seedlings
  • Tandem mass spectrometry
  • Ultraviolet rays
  • Journal article

Cite this

Ge, Y., Cai, Y-M., Bonneau, L., Rotari, V., Danon, A., McKenzie, E. A., ... Gallois, P. (2016). Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis. Cell Death & Differentiation, 23(9), 1493-1501. https://doi.org/10.1038/cdd.2016.34
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. In: Cell Death & Differentiation. 2016 ; Vol. 23, No. 9. pp. 1493-1501.
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abstract = "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 (H2O2, 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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Ge, Y, Cai, Y-M, Bonneau, L, Rotari, V, Danon, A, McKenzie, EA, McLellan, H, Mach, L & Gallois, P 2016, 'Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis', Cell Death & Differentiation, vol. 23, no. 9, pp. 1493-1501. https://doi.org/10.1038/cdd.2016.34

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. (Lead / Corresponding author).

In: Cell Death & Differentiation, Vol. 23, No. 9, 01.09.2016, p. 1493-1501.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis

AU - Ge, Y.

AU - Cai, Y.-M.

AU - Bonneau, L.

AU - Rotari, V.

AU - Danon, A.

AU - McKenzie, E. A.

AU - McLellan, H.

AU - Mach, L.

AU - Gallois, P.

N1 - The Bioimaging Facility microscopes used in this study were purchased with grants from BBSRC, Wellcome Trust and the University of Manchester Strategic Fund. The project was partially funded by BBSRC Grants 34/P14516 and BB/K009478/1.

PY - 2016/9/1

Y1 - 2016/9/1

N2 - 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 (H2O2, 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.

AB - 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 (H2O2, 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.

KW - Amino acid sequence

KW - Apoptosis

KW - Arabidopsis

KW - Arabidopsis proteins

KW - Caspase inhibitors

KW - Cathepsin B

KW - Chromatography, High Pressure Liquid

KW - Endoplasmic reticulum stress

KW - Hydrogen peroxide

KW - Oxidative stress

KW - Paraquat

KW - Phylogeny

KW - Plants, Genetically modified

KW - Recombinant proteins

KW - Seedlings

KW - Tandem mass spectrometry

KW - Ultraviolet rays

KW - Journal article

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DO - 10.1038/cdd.2016.34

M3 - Article

VL - 23

SP - 1493

EP - 1501

JO - Cell Death & Differentiation

JF - Cell Death & Differentiation

SN - 1350-9047

IS - 9

ER -