权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

AT SPECIFIC DNA BINDING PROPERTIES OF A HUMAN PROTEIN

AT 人类蛋白质的特定 DNA 结合特性

基本信息

批准号：
6179364
负责人：
RAYMOND REEVES
金额：
$ 21.97万
依托单位：
WASHINGTON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-08-01 至 2002-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6179364
关键词：
DNA footprinting chemical binding complementary DNA cytokine receptors gene induction /repression genetic promoter element human genetic material tag interleukin 2 intermolecular interaction nonhistone nucleoprotein nucleosomes polymerase chain reaction site directed mutagenesis tissue /cell culture

项目摘要

DESCRIPTION: The HMG-I(Y) group of mammalian "high mobility group" (HMG) chromatin proteins are in vivo regulators of gene expression and founding members of a new class of nonhistone proteins called 'architectural transcription factors'. The investigator has demonstrated in human T cells that HMG-I(Y) proteins participate, along with transcription factors NF-kB, Elf-1 and SRF, in the initial in vivo transcription induction of the interleukin-2 receptor a-chain gene (IL-2Ra), an essential controlling step in mounting an effective immune reaction. The goals of the proposed research are to elucidate, at the molecular/mechanistic level, how different regions of the HMG-I(Y) protein function in the transcriptional regulation of the IL-2Ra gene promoter. The investigator has produced a large number of both variant HMG-I(Y) proteins and mutant IL-2Ra promoter DNAs for use in both in vitro and in vivo investigations of the protein-DNA complexes formed on the IL-2Ra promoter during transcriptional activation. The Specific Aims of the research are to use these mutant proteins and promoter DNAs to: 1) Determine the relative importance of different peptide domains of the HMG-I(Y) protein in the formation of a looped, stereospecific activation complex on the promoter of the human IL-2Ra gene and investigate the molecular events involved in such complex formation in vitro and in vivo. 2) Determine the effects of site-specific HMG-I(Y) protein phosphorylations on protein-protein and protein-DNA interactions during promoter complex formation in vitro and in vivo. 3) Investigate the role played by HMG-I(Y) protein-induced alterations of the nucleosomal chromatin structure of the IL-2Ra promoter region during gene transcriptional activation in vitro and in vivo. A variety of extremely sensitive and quantitative biochemical, biophysical, and biological techniques will be employed to analyze the ability of mutant HMG-I(Y) proteins to participate in IL-2Ra promoter complex formation and function. Results from these experiments will contribute significant new information concerning the regulated expression of the IL-2Ra gene, one of the rate limiting steps in T cell activation during induction of the immune response.

描述：哺乳动物“高迁移率群”(HMG) 的 HMG-I(Y) 群染色质蛋白是基因表达和建立的体内调节因子一类新的非组蛋白的成员，称为“建筑蛋白” 转录因子'。研究人员已经在人类 T 细胞中证明 HMG-I(Y) 蛋白与转录因子 NF-kB 一起参与， Elf-1和SRF，在最初的体内转录诱导中白介素 2 受体 a 链基因 (IL-2Ra)，一个重要的控制步骤建立有效的免疫反应。拟议的目标研究的目的是在分子/机制水平上阐明有何不同 HMG-I(Y) 蛋白区域在转录调控中发挥作用 IL-2Ra基因启动子的。调查者已经制作了大量的变体 HMG-I(Y) 蛋白和突变体 IL-2Ra 启动子 DNA 用于对形成的蛋白质-DNA 复合物进行体外和体内研究转录激活过程中 IL-2Ra 启动子的作用。具体目标研究的主要目的是利用这些突变蛋白和启动子 DNA 来：1) 确定不同肽域的相对重要性 HMG-I(Y) 蛋白在环状、立体特异性激活中的形成人 IL-2Ra 基因启动子上的复合物并研究分子事件涉及体外和体内这种复合物的形成。 2) 确定位点特异性 HMG-I(Y) 蛋白磷酸化的影响启动子复合体中蛋白质-蛋白质和蛋白质-DNA相互作用的研究体外和体内的形成。 3) 调查HMG-I(Y)所扮演的角色蛋白质诱导的核小体染色质结构的改变体外基因转录激活过程中的 IL-2Ra 启动子区域体内。多种极其灵敏定量的生化、将采用生物物理和生物技术来分析突变HMG-I(Y)蛋白参与IL-2Ra启动子的能力复杂的形成和功能。这些实验的结果将提供有关调控表达的重要新信息 IL-2Ra 基因，T 细胞激活的限速步骤之一在诱导免疫反应期间。