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

Molecular analysis of TEF-1 family transcription factors involved in cardiovascular development and function

参与心血管发育和功能的 TEF-1 家族转录因子的分子分析

基本信息

批准号：
12835010
负责人：
OHKUBO Hiroaki
金额：
$ 2.3万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12835010/
关键词：
TEF-1 transcription factor Scalloped Vestigial mVRF Tondu ETF Tead-2 gene Enhancer Tead-2遺伝子

项目摘要

The TEF-1 family of transcription factors consists of at least four members, Tead1/TEF-1, Tead2/ETF, Tead3/ETFR-1 and Tead4/ETFR-2, containing a highly conserved TEA DNA-binding domain, and have been implicated in cardiovascular development and function. It has recently been reported that the scalloped (Sd) protein, a Drosophila homologue of TEF-1, controls the gene expression involved in the wing development through protein-protein interaction with Vestigial (Vg). We have isolated a mouse cDNA encoding amino acid sequences similar to those of the Sd-binding domain (SdBD) of Vg by RTPCR and RACE methods. The protein deduced from the cDNA, termed mouse Vg-related Factor (mVRF), consists of 307 amino acid residues. The amino acid sequence of mVRF showed 49 % identity to that of Tondu, a recently identified human counterpart of Vg, but diverged from that of Drosophila Vg except for that of SdBD. Northern blot analysis revealed that mVRF mRNA expressed in placenta, embryo and testis. Transient transfection assay using the luciferase reporter gene and mVRF expression vector suggested that overexpression of mVRF represses the TEF-1-dependent transcriptional activity. On the other hand, we have identified and characterized the cell-specific 117-bp enhancer sequence in the first intron of the mouse ETF/Tead2 gene required for transcriptional activation in ETF/Tead2 gene expressing cells, such as P19 cells. We have also identified one GC-rich sequence (the GC box) and two tandemly repeated GA-rich sequences (the proximal and distal GA elements) in the enhancer and characterized their possible binding proteins, which may play a role in the cell-specific enhancer function.

TEF-1转录因子家族由至少四个成员Tead1/TEF-1、Tead2/ETF、Tead3/ETFR-1和Tead4/ETFR-2组成，含有一个高度保守的TEA DNA结合域，与心血管的发育和功能有关。最近有报道称，扇贝蛋白(Scaloped，SD)是一种与TEF-1同源的果蝇蛋白，它通过与残留蛋白(Vg)的蛋白相互作用来调控与翅膀发育相关的基因表达。我们用RT-PCR和RACE方法克隆了一个与VG的SD结合区(SdBD)相似的编码氨基酸序列的小鼠cDNA。该基因编码的蛋白质称为小鼠VG相关因子(MVRF)，由307个氨基酸残基组成。MVRF的氨基酸序列与新近发现的人类VG的同源性为49%，而与果蝇VG的氨基酸序列除SdBD外均有差异。Northern印迹分析表明，mVRF在胎盘、胚胎和睾丸中均有表达。利用荧光素酶报告基因和mVRF表达载体的瞬时转染实验表明，mVRF的过表达抑制了TEF-1依赖的转录活性。另一方面，我们已经鉴定并鉴定了小鼠ETF/Tead2基因第一内含子中细胞特异性的117-bp增强子序列，这是表达ETF/Tead2基因的细胞(如P19细胞)转录激活所必需的。我们还在增强子中鉴定了一个富含GC的序列(GC盒)和两个重复的富含GA的序列(近端和远端的GA元件)，并鉴定了它们可能的结合蛋白，这可能在细胞特异性增强子功能中发挥作用。