RNA BINDING PROTEINS IN EPILEPSY AND NEUROLOGIC DISEASE

描述(摘自申请者的摘要)：这是一项竞争性续展 建议拨款研究新型干基蛋白及其在 癫痫。脑干基因缺陷的小鼠表现为癫痫发作 我们的工作表明，与其突变表型一致，肉干 在中枢神经系统的神经元中转录水平相对较高 而这个杰基结合了信使核糖核酸。我们还表明，识别杰基的抗体是 某些自身免疫性神经元病患者的血清中存在 变性(副肿瘤性疾病，PND)。其他研究表明， 人类Jerky基因是儿童失神癫痫(CAE)的候选基因。我们现在 把杰基理解为进化上保守的 包含一个新的RNA结合基序的RNA结合蛋白(RNP)家族。 RNPs是介导mRNAs转录后加工的反式作用因子 和前mRNAs，包括剪接、聚腺苷酸化、转运、靶向、 稳定性和平移性。我们假设在突变小鼠中缺少干酪 导致某些损害神经元的mRNAs的处理存在缺陷 导致癫痫发作的功能。我们还表明，缺乏FMRP(脆性X 智力低下蛋白)，另一种RNP，其失活导致脆性X 综合征，并被认为参与了mRNA的加工，也导致了 在老鼠的癫痫发作中。这一发现与该病的高发病率相一致 脆性X患者的癫痫发作。由于缺乏FMRP的动物代表着第二个 例如，RNP中的异常导致癫痫发作，我们 提示RNP功能障碍可能是癫痫更普遍的发病机制。 由于RNPs在癫痫中的潜在重要性，我们目前的重点是 GRANT的应用是研究干基、类干蛋白的细胞作用， 和FMRP。我们建议：1)分析人类的RNA结合特性 牛肉干蛋白和类似的人类蛋白HHJRK，II)以鉴定细胞 JUKY和FMRP的结合靶点(用我们最近开发的一种方法 实验室)，并为这些目标分配职能，以及3)雇用杰基 自身抗体作为研究Jerky-RNA复合体的工具。这些拟议的实验 将把牛肉干家族建立为具有新的RNA的一组不同的RNP 有约束力的主题。此外，为JUKY和FMRP指定目标将允许我们 将这些靶标连接到细胞通路，并确定这些通路是如何 有助于这些蛋白质的整体功能。最后，这些 实验将有助于我们理解生物多样性的某些方面 癫痫和自身免疫性疾病的发病机制。

DESCRIPTION (from applicant's abstract): This is a competing continuation proposal of a grant funded to study the novel Jerky protein and its role in epilepsy. The mouse line defective in the jerky gene shows epileptic seizures and our work has shown that consistent with its mutant phenotype, jerky is transcribed at a relatively high level in neurons of the central nervous system and that Jerky binds mRNA. We also showed that antibodies recognizing Jerky are present in sera of patients suffering of a certain from of autoimmune neuronal degeneration (paraneoplastic disorders, PND). Other studies suggested that the human jerky gene is a candidate for childhood absence epilepsy (CAE). We now understand Jerky to be a prototypic member of an evolutionarily conserved family of RNA binding proteins (RNPs) containing a novel RNA binding motif. RNPs are trans-acting factors mediating posttranscriptional processing of mRNAs and pre-mRNAs, including splicing, polyadenylation, transport, targeting, stability and translation. We hypothesize that lack of Jerky in mutant mice leads to a deficiency in the processing of certain mRNAs compromising neuronal functions that results in seizures. We also show that lack of FMRP (Fragile X Mental Retardation Protein), another RNP whose inactivation causes fragile X syndrome and which is believed to be involved in mRNA processing, also results in seizures in mice. This finding is consistent with the high incidence of seizures in fragile X patients. Since FMRP-deficient animals represent a second example of a situation in which abnormalities in an RNP result in seizures, we suggest that RNP dysfunction may be more general disease mechanism in epilepsy. Due to the potential importance of RNPs in epilepsy, the focus of our current grant application is to study the cellular role of Jerky, Jerky-like proteins, and FMRP. We propose I) to analyze the RNA binding properties of the human JERKY protein and a similar human protein HHJRK, II) to identify the cellular binding targets of JERKY and FMRP (by a method recently developed in our laboratory) and to assign functions for these targets, and 3) to employ Jerky autoantibodies as tool to study Jerky-RNA complexes. These proposed experiments will establish the jerky family as a distinct group of RNPs with a novel RNA binding motif. Also, specifying targets for JERKY and FMRP will allow us to link these targets to cellular pathways and ascertain how these pathways contribute to the overall function of these proteins. Finally, these experiments will aid in our understanding of certain aspects of the pathogenesis of epilepsy and autoimmune diseases.