Department of Biochemistry and Biomedical Sciences, WCU neurocytomics, College of Medicine, Seoul National University, Seoul, Korea.
Cell Death Differ. 2013 Apr;20(4):639-48. doi: 10.1038/cdd.2012.162. Epub 2013 Jan 11.
Promyelocytic leukemia (PML) is a major component of macromolecular multiprotein complexes called PML nuclear-bodies (PML-NBs). These PML-NBs recruit numerous proteins including CBP, p53 and HIPK2 in response to DNA damage, senescence and apoptosis. In this study, we investigated the effect of presenilin (PS), the main component of the γ-secretase complex, in PML/p53 expression and downstream consequences during DNA damage-induced cell death using camptothecin (CPT). We found that the loss of PS in PS knockout (KO) MEFs (mouse embryonic fibroblasts) results in severely blunted PML expression and attenuated cell death upon CPT exposure, a phenotype that is fully reversed by re-expression of PS1 in PS KO cells and recapitulated by γ-secretase inhibitors in hPS1 MEFs. Interestingly, the γ-secretase cleavage product, APP intracellular domain (AICD), together with Fe65-induced PML expression at the protein and transcriptional levels in PS KO cells. PML and p53 reciprocally positively regulated each other during CPT-induced DNA damage, both of which were dependent on PS. Finally, elevated levels of PML-NB, PML protein and PML mRNA were detected in the brain tissues from Alzheimer's disease (AD) patients, where γ-secretase activity is essential for pathogenesis. Our data provide for the first time, a critical role of the PS/AICD-PML/p53 pathway in DNA damage-induced apoptosis, and implicate this pathway in AD pathogenesis.
早幼粒细胞白血病(PML)是一种称为 PML 核体(PML-NBs)的大分子多蛋白复合物的主要成分。这些 PML-NBs 响应 DNA 损伤、衰老和凋亡,招募包括 CBP、p53 和 HIPK2 在内的许多蛋白质。在这项研究中,我们使用喜树碱(CPT)研究了早老素(PS)(γ-分泌酶复合物的主要成分)在 PML/p53 表达和 DNA 损伤诱导的细胞死亡下游后果中的作用。我们发现 PS 敲除(KO)MEFs(小鼠胚胎成纤维细胞)中 PS 的缺失导致 PML 表达严重减弱,并且在 CPT 暴露时细胞死亡减弱,这种表型可以通过 PS1 在 PS KO 细胞中的重新表达完全逆转,并通过 hPS1 MEFs 中的 γ-分泌酶抑制剂再现。有趣的是,γ-分泌酶切割产物 APP 细胞内结构域(AICD)与 Fe65 一起诱导 PS KO 细胞中的 PML 表达在蛋白质和转录水平上。在 CPT 诱导的 DNA 损伤期间,PML 和 p53 相互正向调节彼此,这两者都依赖于 PS。最后,在阿尔茨海默病(AD)患者的脑组织中检测到 PML-NB、PML 蛋白和 PML mRNA 的水平升高,其中 γ-分泌酶活性对发病机制至关重要。我们的数据首次提供了 PS/AICD-PML/p53 途径在 DNA 损伤诱导的细胞凋亡中的关键作用,并暗示该途径参与 AD 的发病机制。