TRANSCRIPTIONAL REGULATION BY HOXA7 IN EMBRYOGENESIS

HOXA7 在胚胎发生中的转录调控

• 批准号: 2403536
• 负责人: Cory Abate-Shen
• 金额: $ 20.85万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2403536
• 关键词: DNA binding protein; affinity chromatography; developmental genetics; embryo /fetus cell /tissue; embryogenesis; gene expression; genetic regulatory element; genetic transcription; homeobox genes; immunocytochemistry; immunoprecipitation; laboratory mouse; molecular cloning; polymerase chain reaction; protein structure function; transcription factor; transfection; western blottings

The studies described in this proposal will characterize the function of HoxA7 in transcriptional regulation during murine embryogenesis. Transcriptional regulatory proteins play a pivotal role in embryogenesis by controlling the expression of phenotypic target genes therefore their, characterization is crucial for understanding the molecular processes that control mammalian development. The hox genes, are master regulators of murine embryogenesis that encode putative transcriptional regulatory proteins. Many elegant biological studies have elucidated the fundamental role of hox genes during development by detailing their coordinate expression, essential function, and mode of regulation. However, many issues remained unanswered regarding the molecular mechanisms by which Hox proteins exert their action as transcriptional regulators during murine embryogenesis. The goal of this proposal is to characterize the transcriptional properties of a prototype member of the Hox family. HoxA7. HoxA7 provides an appropriate model for this analysis since it has served at a prototype for many of the original studies which have examined the expression and function of hox genes during development and since its primary sequence contains features that are reminiscent of other key transcriptional regulatory proteins. Our strategy will be to perform a detailed biochemical analysis couched within the framework of relevant biochemical systems. Our specific plans are: (I) We will characterize the molecular bases of DNA recognition by HoxA7. These studies address the mechanisms by which HoxA7 interacts selectively with DNA regulatory elements and will facilitate subsequent identification of target genes. (II) Our second goal is to characterize the transcriptional regulatory properties of HoxA7. These studies provide the framework for addressing the function of HoxA7 in the variety of distinct cell populations in which is functional during embryogenesis. (IIl) Protein factors that associate with HoxA7 will be identified and their contribution to functional specificity of HoxA7 will be evaluated. Identification of protein partners is an important step towards recapitulating functional HoxA7 protein complexes, and will provide new insight as to HoxA7 function. (IV). Finally. we will characterize the expression of HoxA7 in the developing embryo using specific polyclonal antisera that we have generated. This analysis will provide a biological context for evaluating the functional significance of the genomic target sequences, transcriptional properties, and interacting proteins factors defined in the other specific Aims. Together, these studies provide an integrated approach which will impart real insight into the function of a prototype Hox protein in transcriptional regulation during embryogenesis.

该提案中描述的研究将表征 鼠胚发生过程中的转录调节中的HOXA7。 转录调节蛋白在胚胎发生中起关键作用 通过控制表型靶基因的表达，因此 表征对于理解分子过程至关重要 控制哺乳动物的发展。 HOX基因是主调节器 编码推定的转录调节的鼠类胚胎发生 蛋白质。 许多优雅的生物学研究阐明了 HOX基因在开发过程中的基本作用是通过详细介绍其 协调表达，基本功能和调节方式。 但是，关于分子的许多问题仍未得到答复 HOX蛋白将其作用作为转录的机制 鼠类胚胎发生过程中的调节剂。 该提议的目的是表征转录 HOX家族原型成员的特性。 Hoxa7。 Hoxa7提供 该分析的适当模型，因为它已经在原型 对于许多原始研究，都检查了表达和 HOX基因在开发过程中的功能及其主要序列 包含让人联想到其他关键转录的功能 调节蛋白。 我们的策略将是执行详细的 生化分析在相关生化的框架内 系统。 我们的具体计划是：（i）我们将表征 Hoxa7的DNA识别。 这些研究涉及的机制 HOXA7与DNA调节元件有选择性相互作用，将会 促进随后鉴定靶基因。 （ii）我们的第二个 目的是表征转录调节性能 Hoxa7。 这些研究提供了解决功能的框架 Hoxa7在各种不同的细胞种群中 胚胎发生期间的功能。 （IIL）关联的蛋白质因子 将确定Hoxa7并对功能的贡献 将评估Hoxa7的特异性。 蛋白质的鉴定 合作伙伴是介绍功能性HOXA7的重要一步 蛋白质复合物，并将为HOXA7功能提供新的见解。 （iv）。最后。我们将表征Hoxa7在 使用特定的多克隆抗血清开发胚胎 生成。 该分析将为 评估基因组靶序列的功能意义， 转录特性和相互作用的蛋白质因子定义 另一个具体目标。 这些研究共同提供了一种综合方法，该方法将赋予 真正了解原型HOX蛋白的功能 胚胎发生过程中的转录调节。