Identification of a functional CBF-binding CCAAT-like motif in the core promoter of the mouse pro-α1(V) collagen gene (Col5a1)
Identification of a functional CBF-binding CCAAT-like motif in the core promoter of the mouse pro-α1(V) collagen gene (Col5a1)
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DOI：
10.1016/j.matbio.2004.03.003
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发表时间：
2004-05-01
期刊：
影响因子：
6.9
通讯作者：
Yoshioka, H
中科院分区：
文献类型：
--
作者：
Sakata-Takatani, K;Matsuo, N;Yoshioka, H
我们使用的核心启动子区域的结构-功能分析，以阐明小鼠的一个I（V）胶原蛋白基因（Col 5a 1）的转录特征。通过瞬时转染实验，在紧邻主要转录起始位点上游的23 1bp内确定了核心启动子，其具有典型的TATA基序并且具有高GC含量。在该区域，我们通过电泳迁移率变动分析确定了三个核因子结合位点：BS 1（-195至-167），BS 2（-134至-106）和BS 3（-110至-80）。寡核苷酸竞争和超移位分析显示，Sp1，CBF，和Sp1相关蛋白特异性结合BSI，BS 2，和BS 3，分别。CCAAT样基序、CAAAT和侧翼序列在小鼠和人基因之间是保守的。CBF，它承认这一主题，激活Col 5a 1启动子，如以前报道的Colla 1和Colla 2。此外，CBF-B亚基的野生型和突变形式的过表达改变了这种活性。这些结果表明，CBF是I型和V型胶原基因协调表达的关键因素。(C)2004年Elsevier B. V./国际矩阵生物学学会。All rights reserved.
We used structure-function analysis of the core promoter region to elucidate the transcriptional features of the mouse a I (V) collagen gene (Col5a1). The core promoter, which tacks a typical TATA motif and has a high GC content, was defined within the 23 1 bp immediately upstream from the major transcription start site by transient transfection experiments. In this region, we identified three nuclear-factor binding sites by electrophoretic mobility shift assay: BS1 (- 195 to - 167), BS2 (- 134 to - 106), and BS3 (-110 to -80). Oligonucleotide competition and supershift assays revealed that Sp1, CBF, and Sp1-related protein specifically bind to BSI, BS2, and BS3, respectively. The CCAAT-like motif, CAAAT, and flanking sequences are conserved between the mouse and human gene. CBF, which recognizes this motif, activated the Col5a1 promoter, as previously reported for Colla1 and Colla2. Furthermore, overexpression of a wild-type and mutant forms of CBF-B subunit altered this activity. These results suggest that CBF is a key factor in the coordinated expression of type I and V collagen genes. (C) 2004 Elsevier B.V./International Society of Matrix Biology. All rights reserved.