Challa Madhavi, Malladi Srinivas, Pellock Brett J, Dresnek Douglas, Varadarajan Shankar, Yin Y Whitney, White Kristin, Bratton Shawn B
Division of Pharmacology and Toxicology, College of Pharmacy, The University of Texas at Austin, Austin, TX, USA.
EMBO J. 2007 Jul 11;26(13):3144-56. doi: 10.1038/sj.emboj.7601745. Epub 2007 Jun 7.
Although essential in mammals, in flies the importance of mitochondrial outer membrane permeabilization for apoptosis remains highly controversial. Herein, we demonstrate that Drosophila Omi (dOmi), a fly homologue of the serine protease Omi/HtrA2, is a developmentally regulated mitochondrial intermembrane space protein that undergoes processive cleavage, in situ, to generate two distinct inhibitor of apoptosis (IAP) binding motifs. Depending upon the proapoptotic stimulus, mature dOmi is then differentially released into the cytosol, where it binds selectively to the baculovirus IAP repeat 2 (BIR2) domain in Drosophila IAP1 (DIAP1) and displaces the initiator caspase DRONC. This interaction alone, however, is insufficient to promote apoptosis, as dOmi fails to displace the effector caspase DrICE from the BIR1 domain in DIAP1. Rather, dOmi alleviates DIAP1 inhibition of all caspases by proteolytically degrading DIAP1 and induces apoptosis both in cultured cells and in the developing fly eye. In summary, we demonstrate for the first time in flies that mitochondrial permeabilization not only occurs during apoptosis but also results in the release of a bona fide proapoptotic protein.
虽然线粒体外膜通透性在哺乳动物细胞凋亡过程中至关重要,但在果蝇中,其对于凋亡的重要性仍极具争议。在此,我们证明了果蝇Omi(dOmi),即丝氨酸蛋白酶Omi/HtrA2的果蝇同源物,是一种在发育过程中受到调控的线粒体内膜间隙蛋白,它会在原位进行连续切割,产生两个不同的凋亡抑制蛋白(IAP)结合基序。根据促凋亡刺激的不同,成熟的dOmi随后会以不同方式释放到细胞质中,在那里它会选择性地与果蝇IAP1(DIAP1)中的杆状病毒IAP重复序列2(BIR2)结构域结合,并取代起始半胱天冬酶DRONC。然而,仅这种相互作用不足以促进细胞凋亡,因为dOmi无法从DIAP1的BIR1结构域中取代效应半胱天冬酶DrICE。相反,dOmi通过蛋白水解降解DIAP1来减轻DIAP1对所有半胱天冬酶的抑制作用,并在培养细胞和发育中的果蝇眼中诱导细胞凋亡。总之,我们首次在果蝇中证明,线粒体通透性不仅在细胞凋亡过程中发生,而且还会导致一种真正的促凋亡蛋白的释放。