CASH--A NOVEL BHLH PROTEIN--ROLE IN GENE TRANSCRIPTION

CASH——一种新型 BHLH 蛋白质——在基因转录中的作用

基本信息

批准号：
2259743
负责人：
ANGELA R PORTA
金额：
$ 5.28万
依托单位：
ROCHE INSTITUTE OF MOLECULAR BIOLOGY
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-09-30 至 1995-08-31
项目状态：
已结题

项目摘要

Proteins regulating gene transcription have been identified, characterized, and classified based on common structural motifs. One motif identified in a growing family of transcription factors is the basic helix-loop-helix (bHLH) domain. Proteins manifesting the BHLH domain participate in the regulation of cell specification and differentiation. For example proteins from the Drosophila achaete-scute complex, regulate neuronal development. A CDNA clone encoding a novel member of the BHLH protein superfamily homologous to these proteins has been isolated from a catfish (Ictalurus punctatus) olfactory epithelium CDNA library. This BHLH protein, named CASH (catfish achaete-acute homolog) is particularly interesting as olfactory epithelium contains neuronal precursors capable of continuous division and differentiation to form olfactory neurons. In light of the role that BHLH transcription factors play in the regulation of cell specification and differentiation, this proposal will study and characterize this BHLH protein and the role in plays in regulating gene expression. To achieve this we plan to; -determine the tissue and cell distribution of the CASH MRNA; express the CASH BHLH protein for antiserum production for immunocytochemical localization and functional studies; identify the DNA sequence motif to which the CASH BHLH protein binds; characterize homo- and heteromeric protein-protein interactions of the CASH protein; evaluate the role of post-translational modifications on the activities of this proteins; identify and characterize the genes that the CASH protein interacts with and evaluate how they are regulated; determine if there are other related BHLH proteins in olfactory mucosa of teleosts and other vertebrates. This will expand knowledge of the BHLH family of transcription factors and advance our understanding of the molecular basis by which these proteins regulate gene transcription. Detailed knowledge of BHLH proteins and the normal mechanisms by which they regulate gene transcription is critical in understanding how disturbances in gene transcription result in metabolic and degenerative diseases associated with aging, oncogenesis, and in response to trauma. This research, coupled with coursework in neuroscience and molecular biology, will upgrade the applicant's skills and serve as an essential bridge for her process and development as an independent investigator so that she may eventually obtain an academic position to pursue independent research studying gene regulation in the nervous system.

调节基因转录的蛋白质已被鉴定，根据常见的结构主题进行表征和分类。一在一个不断增长的转录因子家族中发现的基序是基本螺旋-环-螺旋 (bHLH) 结构域。表现出 BHLH 的蛋白质域参与细胞规范的调节和差异化。例如来自果蝇 achaete-scute 的蛋白质复杂，调节神经元发育。编码小说的 cDNA 克隆与这些蛋白同源的 BHLH 蛋白超家族成员从鲶鱼（Ictalurus punctatus）嗅上皮中分离出来 cDNA 文库。这种BHLH蛋白被命名为CASH（鲶鱼acaete-acute 同系物）特别有趣，因为嗅觉上皮含有能够连续分裂和分化的神经元前体形成嗅觉神经元。鉴于 BHLH 转录的作用因素在细胞规范和分化的调节中发挥作用，该提案将研究和表征这种 BHLH 蛋白及其作用参与调节基因表达。为了实现这一目标，我们计划： -确定CASH mRNA的组织和细胞分布；表达用于免疫细胞化学抗血清生产的 CASH BHLH 蛋白定位和功能研究；识别 DNA 序列基序 CASH BHLH 蛋白结合；表征同聚体和异聚体 CASH 蛋白的蛋白质-蛋白质相互作用；评估的作用该蛋白质活性的翻译后修饰；识别并表征与 CASH 蛋白相互作用的基因并评估它们的监管方式；判断是否还有其他相关硬骨鱼和其他脊椎动物嗅粘膜中的 BHLH 蛋白。这将扩展对 BHLH 转录因子家族的了解并增进我们对这些物质的分子基础的理解蛋白质调节基因转录。 BHLH详细知识蛋白质及其调节基因的正常机制转录对于理解基因中的干扰如何至关重要转录导致代谢和退行性疾病相关与衰老、肿瘤发生以及对创伤的反应有关。这项研究，加上神经科学和分子生物学的课程，将提升申请人的技能并成为她的重要桥梁作为一名独立调查员的过程和发展，以便她可以最终获得学术职位以进行独立研究研究神经系统中的基因调控。