Starfish Apaf-1 activates effector caspase-3/9 upon apoptosis of aged eggs

Ritsuko Tamura, Mariko Takada, Miki Sakaue, Ayaka Yoshida, Shirabe Ohi, Kaoru Hirano, Tomoyo Hayakawa, Noritaka Hirohashi, Kei Yura, Kazuyoshi Chiba

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Caspase-3-related DEVDase activity is initiated upon apoptosis in unfertilized starfish eggs. In this study, we cloned a starfish procaspase-3 corresponding to mammalian effector caspase containing a CARD that is similar to the amino terminal CARD of mammalian capsase-9, and we named it procaspase-3/9. Recombinant procaspase-3/9 expressed at 15 °C was cleaved to form active caspase-3/9 which has DEVDase activity. Microinjection of the active caspase-3/9 into starfish oocytes/eggs induced apoptosis. An antibody against the recombinant protein recognized endogenous procaspase-3/9 in starfish oocytes, which was cleaved upon apoptosis in aged unfertilized eggs. These results indicate that caspase-3/9 is an effector caspase in starfish. To verify the mechanism of caspase-3/9 activation, we cloned starfish Apaf-1 containing a CARD, a NOD, and 11 WD40 repeat regions, and we named it sfApaf-1. Recombinant sfApaf-1 CARD interacts with recombinant caspase-3/9 CARD and with endogenous procaspase-3/9 in cell-free preparations made from starfish oocytes, causing the formation of active caspase-3/9. When the cell-free preparation without mitochondria was incubated with inactive recombinant procaspase-3/9 expressed at 37 °C, DEVDase activity increased and apoptosome-like complexes were formed in the high molecular weight fractions containing both sfApaf-1 and cleaved caspase-3/9. These results suggest that sfApaf-1 activation is not dependent on cytochrome c.

Original languageEnglish
Article number1611
JournalScientific Reports
Volume8
Issue number1
DOIs
Publication statusPublished - 2018 Dec 1
Externally publishedYes

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Effector Caspases
Starfish
Caspase 9
Caspase 3
Eggs
Apoptosis
Oocytes
Ovum
Apoptosomes
Microinjections

ASJC Scopus subject areas

  • General

Cite this

Tamura, R., Takada, M., Sakaue, M., Yoshida, A., Ohi, S., Hirano, K., ... Chiba, K. (2018). Starfish Apaf-1 activates effector caspase-3/9 upon apoptosis of aged eggs. Scientific Reports, 8(1), [1611]. https://doi.org/10.1038/s41598-018-19845-6

Starfish Apaf-1 activates effector caspase-3/9 upon apoptosis of aged eggs. / Tamura, Ritsuko; Takada, Mariko; Sakaue, Miki; Yoshida, Ayaka; Ohi, Shirabe; Hirano, Kaoru; Hayakawa, Tomoyo; Hirohashi, Noritaka; Yura, Kei; Chiba, Kazuyoshi.

In: Scientific Reports, Vol. 8, No. 1, 1611, 01.12.2018.

Research output: Contribution to journalArticle

Tamura, R, Takada, M, Sakaue, M, Yoshida, A, Ohi, S, Hirano, K, Hayakawa, T, Hirohashi, N, Yura, K & Chiba, K 2018, 'Starfish Apaf-1 activates effector caspase-3/9 upon apoptosis of aged eggs', Scientific Reports, vol. 8, no. 1, 1611. https://doi.org/10.1038/s41598-018-19845-6
Tamura R, Takada M, Sakaue M, Yoshida A, Ohi S, Hirano K et al. Starfish Apaf-1 activates effector caspase-3/9 upon apoptosis of aged eggs. Scientific Reports. 2018 Dec 1;8(1). 1611. https://doi.org/10.1038/s41598-018-19845-6
Tamura, Ritsuko ; Takada, Mariko ; Sakaue, Miki ; Yoshida, Ayaka ; Ohi, Shirabe ; Hirano, Kaoru ; Hayakawa, Tomoyo ; Hirohashi, Noritaka ; Yura, Kei ; Chiba, Kazuyoshi. / Starfish Apaf-1 activates effector caspase-3/9 upon apoptosis of aged eggs. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
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