Kovacs D M, Mancini R, Henderson J, Na S J, Schmidt S D, Kim T W, Tanzi R E
Department of Neurology, Massachusetts General Hospital East, Harvard Medical School, Charlestown 02129, USA.
J Neurochem. 1999 Dec;73(6):2278-85. doi: 10.1046/j.1471-4159.1999.0732278.x.
Familial Alzheimer's disease (FAD) mutant forms of presenilin 1 (PS1) and 2 have been shown to sensitize cells to apoptotic cell death. Here we explore the effects of FAD mutant forms of PS1 on caspase activation during apoptosis. We show that caspase activation leads to increased generation of alternative C-terminal fragments (CTFs) from mutant as compared to wild-type (wt) PS1. For this purpose, very low expression levels of wt, A246E, L286V, and deltaE10 FAD mutant PS1 proteins in stably transfected human H4 neuroglioma cells were used to avoid artifactual induction of spontaneous apoptosis due to overexpression of PS1. Staurosporine treatment of these cells resulted in increased cell death and up to a 10-fold increase in caspase-3 activation in mutant versus wt PS1-expressing cell lines. Correspondingly, relative levels of caspase-cleaved PS1 CTFs were increased by five- to sixfold in the FAD mutant versus wt PS1 cells. Elevated caspase activation and caspase cleavage of FAD mutant PS1 suggest the possibility of either a direct proapoptotic effect of mutant PS1 or interference of mutant PS1 with antiapoptotic effects of wt PS1.
早老素1(PS1)和早老素2的家族性阿尔茨海默病(FAD)突变形式已被证明会使细胞对凋亡性细胞死亡敏感。在此,我们探讨FAD突变形式的PS1在细胞凋亡过程中对半胱天冬酶激活的影响。我们发现,与野生型(wt)PS1相比,半胱天冬酶激活会导致突变体产生更多的替代性C末端片段(CTF)。为此,在稳定转染的人H4神经胶质瘤细胞中使用极低表达水平的wt、A246E、L286V和deltaE10 FAD突变型PS1蛋白,以避免因PS1过表达而人为诱导自发凋亡。用星形孢菌素处理这些细胞会导致细胞死亡增加,在表达突变体PS1的细胞系中,半胱天冬酶-3的激活比表达wt PS1的细胞系增加了10倍。相应地,在FAD突变体细胞中,半胱天冬酶切割的PS1 CTF的相对水平比wt PS1细胞增加了五到六倍。FAD突变体PS1的半胱天冬酶激活和切割增加,提示突变体PS1可能具有直接的促凋亡作用,或者干扰了wt PS1的抗凋亡作用。