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*

*この研究の対応する著者

研究成果: Article査読

13 被引用数 (Scopus)

抄録

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.

本文言語English
論文番号1611
ジャーナルScientific reports
8
1
DOI
出版ステータスPublished - 2018 12月 1
外部発表はい

ASJC Scopus subject areas

  • 一般

フィンガープリント

「Starfish Apaf-1 activates effector caspase-3/9 upon apoptosis of aged eggs」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル