HEL-N1 IN NEURONAL RNA PROCESSING AND AUTO IMMUNITY

金博士在这次培训期间的长期目标是开发一种 专注于基础科学方法的研究生涯 了解神经病学的临床疾病。在这两年中 在分子生物学方面，金博士已经建立了他的 尽早致力于实现这一目标。他成功地学到了基础知识 并成功地克隆了一种新的编码一种 神经元特异性RNA结合蛋白Hel-N1。初步研究 表明该蛋白与某些基因的3‘端非翻译区结合 癌基因转录本，即c-myc和c-fos。作为绑定的区域 已经证明在mRNA中参与了转录本 稳定，在Hel-N1和 这些基因的转录后调控。金博士计划 使用信使核糖核酸稳定性分析在他的提案中研究这种可能性 通过对重组Hel-N1病毒进行进一步的结合研究 更准确地描述蛋白质与RNA的相互作用。这些 调查将要求他学习更多的高级方法 包括转基因、诱变和组织培养技术。 有趣的是，Hel-Hel-2的氨基酸序列具有很高的同源性。 N1和几种与副肿瘤、自身免疫综合征有关的抗原 中枢神经系统(CNS)。最常与小蜂窝有关 肺癌，这些综合征涉及免疫调节 破坏中枢神经系统内的各种基因，包括背根 神经节、海马体、小脑和脊髓。金博士将 研究氨基酸序列相似性的意义和 在这些抗原的免疫学方面的差异和 它们产生了不同的临床症状。这将涉及到 使用这些综合征患者的血清进行表位分析。 此外，他还将测试Hel-N1引发中枢神经系统疾病的能力 易感大鼠品系的自身免疫反应 用重组、纯化的蛋白进行免疫。这个动物模型 将有助于更详细地了解这些综合征的机制 S。 总而言之，这项提案的资金将为金博士提供 这是研究Hel-N1病毒功能的绝佳机会 科学和临床水平。这也会让他有必要的时间 作为一名独立的临床研究人员走向成熟。

Dr. King's long term goal during this training period is to develop a research career that focuses on basic science approaches to understanding clinical disease in neurology. During the two year fellowship in molecular biology, Dr. King has already established his early commitment to that goal. He successfully learned the fundamentals of molecular biology and was able to clone a novel cDNA encoding a neuronal-specific RNA binding protein named Hel-N1. Preliminary studies indicate that this protein binds the 3' untranslated regions of certain oncogene transcripts, namely c-myc and c-fos. As the region of binding within the mRNA has already been shown to be involved in transcript stability, there is potentially an exciting link between Hel-N1 and the post-transcriptional regulation of these genes. Dr. King plans to investigate this possibility in his proposal using mRNA stability assays with recombinant Hel-N1 and by performing further binding studies to characterize more precisely the protein-RNA interaction. These investigations will require that he learn more advance approaches including transfections, mutagenesis and tissue culture techniques. Interestingly, there is close amino acid sequence homology between Hel- N1 and several antigens involved in paraneoplastic, autoimmune syndromes of the central nervous system (CNS). Most often related to small cell carcinoma of the lung, these syndromes involve immune mediated destruction of various loci within the CNS, including dorsal root ganglia, hippocampus, cerebellum and spinal cord. Dr. King will investigate the significance of the amino acid sequence similarities and differences with respect to the immunology of these antigens and the heterogeneous clinical syndromes they produce. This will involve epitope analysis using sera from patients with these syndromes. Moreover, he will also test the ability of Hel-N1 to induce a CNS autoimmune response in susceptible rat strains through the use of immunizations with recombinant, purified protein. This animal model will allow for more detailed understanding of the mechanism of these syndro s. In summary, the funding of this proposal will provide Dr. King an excellent opportunity to study the function of Hel-N1 both on a basic science and clinical level. It will also allow him the time necessary to mature as an independent clinical investigator.