In this study, we employed chromatin immunoprecipitation, a useful method for studying the locations of transcription factors bound to specific DNA regions in specific cells, to investigate amyloid precursor protein i...In this study, we employed chromatin immunoprecipitation, a useful method for studying the locations of transcription factors bound to specific DNA regions in specific cells, to investigate amyloid precursor protein intracellular domain binding sites in chromatin DNA from hippocampal neurons of rats, and to screen out five putative genes associated with the learning and memory functions. The promoter regions of the calcium/calmodulin-dependent protein kinase II alpha and glutamate receptor-2 genes were amplified by PCR from DNA products immunoprecipitated by amyloid precursor protein intracellular domain. An electrophoretic mobility shift assay and western blot analysis suggested that the promoter regions of these two genes associated with learning and memory were bound by amyloid precursor protein intracellular domain （in complex form）. Our experimental findings indicate that the amyloid precursor protein intracellular domain is involved in the transcriptional regulation of learning- and memory-associated genes in hippocampal neurons. These data may provide new insights into the molecular mechanism underlying the symptoms of progressive memory loss in Alzheimer＇s disease.展开更多
Amyloid precursor protein intracellular domain （AICD） is one of the potential candidates in deciphering the complexity of Alzheimer＇s disease. It plays important roles in determining cell fate and neurodegeneration...Amyloid precursor protein intracellular domain （AICD） is one of the potential candidates in deciphering the complexity of Alzheimer＇s disease. It plays important roles in determining cell fate and neurodegeneration through its interactions with several adaptors. The pres- ence or absence of phosphorylation at specific sites determines the choice of partners. In this study, we identified 20 novel AICD- interacting proteins by in vitro pull down experiments followed by 2D gel electrophoresis and MALDI-MS analysis. The identified proteins can be grouped into different functional classes including molecular chaperones, structural proteins, signaling and transport molecules, adaptors, motor proteins and apoptosis determinants. Interactions of nine proteins were further validated either by colocal- ization using confocal imaging or by co-immunoprecipitation followed by immunoblotting. The cellular functions of most of the proteins can be correlated with AD. Hence, illustration of their interactions with AICD may shed some light on the disease pathophysiology.展开更多
Alzheimer's disease (AD),a degenerative neurological disorder,is the most common form of dementia among older people,whose symptoms include gradual memory loss,cognitive impairments and deterioration of language s...Alzheimer's disease (AD),a degenerative neurological disorder,is the most common form of dementia among older people,whose symptoms include gradual memory loss,cognitive impairments and deterioration of language skills.Amyloid precursor protein (APP) is cleaved by serials of secretases and generates Aβ,sAPPα/β and APP intracellular domain (AICD).Aβ forms amyloid plaques,together with neurofibrillary tangles (NFTs) which is comprised with hyperphosphorylated tau,are hallmarks ofAD.Aβ,especially in its oligomeric form,plays important roles in AD,causing cell death,calcium influx,loss of spines and repression of long-term potentiation (LTP)[1].However,recent studies indicate that in addition to Aβ,other fragments of APP after its cleavage,such as AICD,play essential roles in AD as well.In this article,the function of AICD and its underlying mechanisms will be reviewed.展开更多
基金supported by the Natural Science Foundation of Guangdong Province,China,No.8151051501000004
文摘In this study, we employed chromatin immunoprecipitation, a useful method for studying the locations of transcription factors bound to specific DNA regions in specific cells, to investigate amyloid precursor protein intracellular domain binding sites in chromatin DNA from hippocampal neurons of rats, and to screen out five putative genes associated with the learning and memory functions. The promoter regions of the calcium/calmodulin-dependent protein kinase II alpha and glutamate receptor-2 genes were amplified by PCR from DNA products immunoprecipitated by amyloid precursor protein intracellular domain. An electrophoretic mobility shift assay and western blot analysis suggested that the promoter regions of these two genes associated with learning and memory were bound by amyloid precursor protein intracellular domain （in complex form）. Our experimental findings indicate that the amyloid precursor protein intracellular domain is involved in the transcriptional regulation of learning- and memory-associated genes in hippocampal neurons. These data may provide new insights into the molecular mechanism underlying the symptoms of progressive memory loss in Alzheimer＇s disease.
基金supported by the Structural Proteomics and Genomics of Human Genetic Disorders Project of Department of Atomic Energy
文摘Amyloid precursor protein intracellular domain （AICD） is one of the potential candidates in deciphering the complexity of Alzheimer＇s disease. It plays important roles in determining cell fate and neurodegeneration through its interactions with several adaptors. The pres- ence or absence of phosphorylation at specific sites determines the choice of partners. In this study, we identified 20 novel AICD- interacting proteins by in vitro pull down experiments followed by 2D gel electrophoresis and MALDI-MS analysis. The identified proteins can be grouped into different functional classes including molecular chaperones, structural proteins, signaling and transport molecules, adaptors, motor proteins and apoptosis determinants. Interactions of nine proteins were further validated either by colocal- ization using confocal imaging or by co-immunoprecipitation followed by immunoblotting. The cellular functions of most of the proteins can be correlated with AD. Hence, illustration of their interactions with AICD may shed some light on the disease pathophysiology.
文摘Alzheimer's disease (AD),a degenerative neurological disorder,is the most common form of dementia among older people,whose symptoms include gradual memory loss,cognitive impairments and deterioration of language skills.Amyloid precursor protein (APP) is cleaved by serials of secretases and generates Aβ,sAPPα/β and APP intracellular domain (AICD).Aβ forms amyloid plaques,together with neurofibrillary tangles (NFTs) which is comprised with hyperphosphorylated tau,are hallmarks ofAD.Aβ,especially in its oligomeric form,plays important roles in AD,causing cell death,calcium influx,loss of spines and repression of long-term potentiation (LTP)[1].However,recent studies indicate that in addition to Aβ,other fragments of APP after its cleavage,such as AICD,play essential roles in AD as well.In this article,the function of AICD and its underlying mechanisms will be reviewed.
