Kim Hye-Sun, Kim Eun-Mee, Lee Jean-Pyo, Park Cheol Hyoung, Kim Seonghan, Seo Ji-Heui, Chang Keun-A, Yu Eunah, Jeong Sung-Jin, Chong Young Hae, Suh Yoo-Hun
Department of Pharmacology, College of Medicine, National Creative Research Initiative Center for Alzheimer's Dementia, Seoul National University, Seoul, 110-799, South Korea.
FASEB J. 2003 Oct;17(13):1951-3. doi: 10.1096/fj.03-0106fje. Epub 2003 Aug 15.
The AICD (amyloid precursor protein [APP] intracellular domain) and C31, the caspase-cleaved C-terminal fragment of APP, have been found in the brains of patients with Alzheimer's disease (AD). Here, we demonstrate for the first time that the C-terminal fragments of APP (AICD [C57, C59] and C31) exert neurotoxicity on differentiated PC 12 cells and rat primary cortical neurons by inducing the expression of glycogen synthase kinase 3beta, forming a ternary complex with Fe65 and CP2/LSF/LBP1 in the nucleus, whereas deletion mutants and a point mutant with Y682G of the YENPTY domain, a Fe65 binding domain, do not. Moreover, expression of APP770 and Swedish mutant form of APP increased the levels of C-terminal fragments of APP (APP-CTs) in neuronal cells and also induced the up-regulation of glycogen synthase kinase-3beta at both the mRNA and the protein levels. In addition, we show that CP2/LSF/LBP1 binding site (nt +0 to approximately +10) in human glycogen synthase kinase 3beta promoter region is essential for the induction of the gene transcription by APP-CTs. The neurotoxicities induced by APP-CTs (AICD and C31) were accompanied by an increase in the active form of glycogen synthase kinase-3beta, and by the induction of tau phosphorylation and a reduction in nuclear beta-catenin levels, and led to apoptosis.
淀粉样前体蛋白(APP)细胞内结构域(AICD)以及APP经半胱天冬酶切割后的C末端片段C31,已在阿尔茨海默病(AD)患者的大脑中被发现。在此,我们首次证明APP的C末端片段(AICD [C57、C59]和C31)通过诱导糖原合酶激酶3β的表达,对分化的PC12细胞和大鼠原代皮质神经元产生神经毒性,在细胞核中与Fe65和CP2/LSF/LBP1形成三元复合物,而Fe65结合结构域YENPTY结构域的缺失突变体和Y682G点突变体则不会。此外,APP770和瑞典突变型APP的表达增加了神经元细胞中APP的C末端片段(APP-CTs)水平,并且在mRNA和蛋白质水平上均诱导了糖原合酶激酶-3β的上调。另外,我们表明人糖原合酶激酶3β启动子区域中的CP2/LSF/LBP1结合位点(核苷酸+0至约+10)对于APP-CTs诱导基因转录至关重要。APP-CTs(AICD和C31)诱导的神经毒性伴随着糖原合酶激酶-3β活性形式的增加、tau蛋白磷酸化的诱导以及细胞核β-连环蛋白水平的降低,并导致细胞凋亡。