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Activation of Plp Gene Expression in Oligodendrocytes

少突胶质细胞中 Plp 基因表达的激活

基本信息

批准号：
7932841
负责人：
Patricia A. Wight
金额：
$ 32.34万
依托单位：
UNIV OF ARKANSAS FOR MED SCIS
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-07-15 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7932841
关键词：
Accounting Action Potentials Address Adult Adverse effects Affinity Chromatography Age Axon Binding Brain Cause of Death Cell Line Cell membrane Cells Childhood Cognitive deficits Complex DNA DNA Binding DNase-I Footprinting Data Demyelinating Diseases Demyelinations Development Disease Electrophoretic Mobility Shift Assay Enhancers Etiology Exhibits Future Gene Activation Gene Expression Gene Proteins Genes Genetic Recombination Genetic Transcription Heat shock proteins Human Introns LacZ Genes Lead Link Maps Mass Spectrum Analysis Mediating Messenger RNA Methods Modeling Molecular Monitor Motor Mouse Strains Multiple Sclerosis Mus Mutate Mutation Myelin Myelin Proteolipid Protein Names Neuraxis Neurodegenerative Disorders Neuroglia Nuclear Nuclear Extract Nuclear Protein Nuclear Proteins Nucleoproteins Oligodendroglia Pathologic Patients Pattern Pelizaeus-Merzbacher Disease Peripheral Nervous System Positioning Attribute Pregnancy Procedures Process Progress Reports Proteins Proteolipids Publications Regulation Regulatory Element Relative (related person)Reporter Genes Schwann Cells Sensory Disorders Site Spastic Paraplegia Speed Spinal cord injury Synaptic Transmission Testing Therapeutic Time Transcription Coactivator Transfection Transgenes Transgenic Mice Transgenic Organisms Western Blotting Work axonopathy base cell type chromatin immunoprecipitation design dysmyelination early childhood early onset fusion gene in vivo insight myelination nervous system development new therapeutic target null mutation plasma protein Z postnatal promoter protein complex spatiotemporal therapy design three dimensional structure

项目摘要

The myelin proteolipid protein (PLP) gene (Plp1) encodes the most abundant protein present in CNS myelin. Its expression in oligodendrocytes is tightly regulated. Mutations in the human gene have been shown to be associated with the X-linked dysmyelinating disorder Pelizaeus-Merzbacher disease (PMD) and some types of spastic paraplegia (SPG2). Dysmyelination can arise from either elevated levels of Plp1 gene expression or the lack, thereof. Hence accurate expression of the gene in is critical. With transgenic mice that harbor Plp1- lacZ fusion genes we have shown that Plp1 intron 1 DNA is essential for regulating the dramatic increase in Plp1 gene transcription in oligodendrocytes during the active myelination period of CNS development. The intron accounts for roughly one-half of the Plp1 gene, encompassing over 8 kb of DNA. We have identified a regulatory element within Plp1 intron 1 DNA and named it ASE for antisilencer/enhancer. We believe the ASE participates in the formation of a multi-protein nucleoprotein complex which acts as a potent transcriptional activator called an enhanceosome. Enhanceosomes are a new class of transcriptional regulators that are characterized by a higher-order nucleoprotein complex which functions as a unit. The precise three- dimensional structure permits synergy between factors and is obligatory to elicit gene activation. The intron also contains another enhancer, designated as wmN1, which has been roughly localized to a 1.2 kb segment of DNA. We hypothesize that the dramatic increase in Plp1 gene activity in oligodendrocytes during development is governed by the ASE and wmN1 enhancers and their associated (cognate) DNA-binding factors. Elucidation of the mechanisms controlling Plp1 gene expression will be an important step in the design of therapies aimed at enhancing remyelination since at the time of myelination, 10% of the total mRNA in oligodendrocytes is generated from the Plp1 gene. Developmental differences in factors present early in childhood versus those in adults may explain the rather poor levels of remyelination observed in MS patients. Cognate factors which bind the ASE and wmN1 enhancers may provide new therapeutic targets to enhance remyelination in these patients. As well, critical mutations in the ASE or wmN1 enhancer, or a mutation in a gene which encodes a cognate factor obligatory for enhancer function, would lead to under-expression of the Plp1 gene and consequently may result in a dysmyelinating disorder. In this study, deletion-transfection analysis with Plp1-lacZ constructs will be used to minimally map the wmN1 enhancer within the 1.2 kb segment of Plp1 intron 1 DNA. Transgenic mice will also be generated to test whether the ASE and wmN1 enhancer work independently, or in combination with one another, to significantly increase the levels of Plp1 gene expression during development.

髓鞘蛋白脂质蛋白（PLP）基因（Plp 1）编码CNS髓鞘中最丰富的蛋白质。其在少突胶质细胞中的表达受到严格调控。人类基因的突变已经被证明是与X连锁髓鞘形成障碍相关的Pelizaeus-Merzbacher病（PMD）和某些类型的痉挛性截瘫（SPG 2）。髓鞘形成障碍可以由Plp 1基因表达水平升高或的缺失，其中。因此，基因的准确表达至关重要。用携带Plp 1基因的转基因小鼠， lacZ融合基因，我们已经表明Plp 1内含子1 DNA对于调节细胞凋亡的急剧增加是必不可少的。中枢神经系统发育活跃髓鞘形成期少突胶质细胞中Plp 1基因的转录。的内含子约占Plp 1基因的一半，包含超过8 kb的DNA。我们已经确定了一 Plp 1内含子1 DNA中的调控元件，并将其命名为ASE，即抗沉默子/增强子。我们相信ASE 参与多蛋白核蛋白复合物的形成，该复合物作为有效的转录因子，一种叫做增强体的激活剂。增强体是一类新的转录调节因子，其特征在于作为一个单元起作用的高级核蛋白复合物。准确的三个- 三维结构允许因子之间的协同作用，并且是引发基因激活的必要条件。内含子还含有另一个增强子，命名为wmN 1，其已大致定位于1.2 kb片段的DNA。我们推测，在少突胶质细胞中Plp 1基因活性的急剧增加，发育受ASE和wmN 1增强子及其相关（同源）DNA结合的控制因素阐明Plp 1基因表达的调控机制将是设计的重要一步目的在于增强髓鞘再生的治疗，因为在髓鞘形成时，少突胶质细胞由Plp 1基因产生。早期存在的因素的发展差异，儿童期与成人期相比，可能解释了在MS患者中观察到的髓鞘再生水平相当差。结合ASE和wmN 1增强子的同源因子可能提供新的治疗靶点，以增强这些患者的髓鞘再生。同样，ASE或wmN 1增强子中的关键突变，或编码增强子功能必需的同源因子的基因，将导致 plp 1基因，因此可能导致髓鞘形成障碍。在本研究中，使用Plp 1-lacZ构建体的分析将用于在1.2kb内最低限度地定位wmN 1增强子 Plp 1内含子1 DNA片段。还将产生转基因小鼠，以测试ASE和wmN 1 增强子单独或相互结合发挥作用，显著增加Plp 1的水平基因在发育过程中的表达