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

REGULATION OF MYOSIN HEAVY CHAIN ALPHA GENE BY CAMP

CAMP 对肌球蛋白重链 α 基因的调控

基本信息

批准号：
2750348
负责人：
RADOVAN ZAK
金额：
$ 31.02万
依托单位：
UNIVERSITY OF CHICAGO
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：
1991-08-26 至 1999-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2750348
关键词：
cyclic AMP gene expression gene targeting genetic promoter element genetic regulation genetic transcription genetically modified animals heart contraction hormone regulation /control mechanism laboratory mouse laboratory rat myosins phosphorylation protein isoforms receptor expression tissue /cell culture transcription factor transfection

项目摘要

DESCRIPTION: The application is a continuation of studies on the mechanism involved in the cAMP-induced regulation of alpha MHC gene transcription. Recent work in the investigator's laboratory has demonstrated that in primary cultures of fetal rat heart myocytes the elevation of intra-cellular cAMP results in up-regulation of alpha MHC and down-regulation of beta MHC mRNA expression. Using several deletion constructs in transient transfection assay, the investigator has identified a 31 base-pair fragment, which contains a hybrid motif with overlapping sequence of E-box and MCAT box (EM element) which confers both muscle specific and cAMP-inducible expression of the gene. The protein that recognizes this element appears to be indistinguishable from the previously characterized TEF-1 transcription factor. The applicant's goal, therefore, is (1) to characterize the protein which interacts with the cAMP-inducible element (EM) in the rat alpha MHC gene promoter by way of isolating and characterizing the cDNA encoding the cardiac specific TEF-1 isoform binding to the EM element, (2) clone and characterize the cDNA of a putative partner protein involved in the interaction, (3) study the cAMP-dependent phosphorylation of factors binding to the EM element (4) define the role of phosphorylation in protein factors which bind to DNA and recognize other proteins in the protein-protein interaction, and (5) verify the in vitro studies by in vivo experiments by constructing transgenic mice and by targeted ablation of selected loci by isolating cardiac specific isogenes from 129 S.V. mouse. It is understood that the proposed experiments will further elucidate the role of signaling pathways in the coupling of the hemodynamic load to the compensatory growth of the heart.

描述：该应用程序是对 CAMP诱导α-MHC基因调控的机制抄写。调查员实验室最近的工作是证明在原代培养的胎鼠心肌细胞中细胞内cAMP升高导致α-MHC上调下调β-MHC基因的表达。使用多个删除在瞬时转染实验中，研究人员已经鉴定出一个31个碱基对的片段，其中包含一个与 E-box和MCAT box(EM元件)的重叠序列肌肉特异性和cAMP诱导的基因表达。这个识别这一元素的蛋白质似乎与先前鉴定的TEF-1转录因子。申请人的因此，目标是(1)表征与之相互作用的蛋白质大鼠α-MHC基因启动子中cAMP诱导元件(EM) 心脏编码基因的分离和鉴定方法与EM元件结合的特异性TEF-1亚型，(2)克隆和鉴定一种可能的伙伴蛋白的cdna。相互作用，(3)研究cAMP依赖的因子磷酸化与EM元件(4)的结合定义了磷酸化在与DNA结合并识别细胞中其他蛋白质的蛋白质因子蛋白质-蛋白质相互作用，以及(5)通过In 构建转基因小鼠和靶向接种的活体实验从129例患者中分离心脏特异性同源基因消融选定的基因小白鼠。据了解，拟议的实验将进一步阐明信号通路在细胞周期蛋白偶联中的作用血流动力学负荷对心脏代偿性生长的影响。