Evolutionarily conserved promoter region containing CArG*-like elements is crucial for smooth muscle myosin heavy chain gene expression.
Evolutionarily conserved promoter region containing CArG*-like elements is crucial for smooth muscle myosin heavy chain gene expression.
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含有 CArG* 样元件的进化保守启动子区域对于平滑肌肌球蛋白重链基因表达至关重要。
DOI：
10.1161/01.res.82.5.566
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发表时间：
1998
影响因子：
20.1
通讯作者：
Periasamy,M
中科院分区：
文献类型：
--
作者：
Zilberman,A;Dave,V;Miano,J;Olson,EN;Periasamy,M
近年来，在了解骨骼肌发育方面取得了重大进展。然而，调节平滑肌发育和分化的机制目前尚不清楚。为了更好地了解平滑肌特异性基因表达，我们将研究重点放在平滑肌肌球蛋白重链（SMHC）基因上，这是一种高度特异性的平滑肌细胞分化标志物。本研究的目的是分离和表征小鼠SMHC基因启动子，因为小鼠启动子将特别适合于转基因小鼠体内启动子分析，并将作为将感兴趣的基因靶向平滑肌细胞的工具。我们在此报道小鼠SMHC启动子及其5 '侧翼区域的分离和表征。对启动子2.6 kb部分的DNA序列分析确定了已知转录因子的几个潜在结合位点。平滑肌细胞原代培养中启动子缺失结构的瞬时转染分析表明，- 1208和- 1050 bp之间的区域是SMHC启动子活性最大化的关键区域。小鼠、大鼠和家兔的SMHC启动子序列比较发现，在- 967和- 1208 bp之间存在一个高度保守的区域。该区域包括三个CArG/CArG*样元件、两个SP-1结合位点、一个nf -1样元件、一个Nkx2-5结合位点和一个Elk-1结合位点。凝胶迁移迁移实验和dna酶I足迹分析表明，这三种CArG/CArG*样元素都可以与血管平滑肌细胞的核提取物形成dna -蛋白复合物。与CArG*元件结合的蛋白质可以被血清反应元件或来自心脏α-肌动蛋白基因的真正CArG元件竞争出去。使用血清反应因子（SRF）抗体，我们证明SRF是蛋白质复合物的一部分。此外，我们发现SRF显性阴性突变表达载体共转染可消除SMHC启动子活性，表明SRF蛋白在SMHC基因调控中起关键作用。
—In recent years, significant progress has been made toward understanding skeletal muscle development. However, the mechanisms that regulate smooth muscle development and differentiation are presently unknown. To better understand smooth muscle–specific gene expression, we have focused our studies on the smooth muscle myosin heavy chain (SMHC) gene, a highly specific marker of differentiated smooth muscle cells. The goal of the present study was to isolate and characterize the mouse SMHC gene promoter, since the mouse promoter would be particularly suited for in vivo promoter analyses in transgenic mice and would serve as a tool for targeting genes of interest into smooth muscle cells. We report here the isolation and characterization of the mouse SMHC promoter and its 5′ flanking region. DNA sequence analysis of a 2.6-kb portion of the promoter identified several potential binding sites for known transcription factors. Transient transfection analysis of promoter deletion constructs in primary cultures of smooth muscle cells showed that the region between −1208 and −1050 bp is critical for maximal SMHC promoter activity. A comparison of SMHC promoter sequences from mouse, rat, and rabbit revealed the presence of a highly conserved region located between −967 and −1208 bp. This region includes three CArG/CArG*-like elements, two SP-1 binding sites, a NF-1–like element, an Nkx2–5 binding site, and an Elk-1 binding site. Gel mobility shift assay and DNase I footprinting analyses show that all three CArG/CArG*-like elements can form DNA-protein complexes with nuclear extract from vascular smooth muscle cells. Protein binding to the CArG* elements can be competed out by either serum response element or by an authentic CArG element from the cardiac α-actin gene. Using a serum response factor (SRF) antibody, we demonstrate that SRF is part of the protein complex. In addition, we show that cotransfection with the SRF dominant-negative mutant expression vector abolishes SMHC promoter activity, suggesting that SRF protein plays a critical role in SMHC gene regulation.
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一种新型平滑肌特异性增强剂调节血管平滑肌细胞中平滑肌肌球蛋白重链基因的转录 (*)
DOI：
10.1074/jbc.270.52.30949
发表时间：
1995
期刊：
The Journal of Biological Chemistry
影响因子：
--
作者：
R. Kallmeier;C. Somasundaram;P. Babij
通讯作者：
P. Babij
兔平滑肌肌球蛋白重链基因功能启动子元件的鉴定。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Katoh,Y;Loukianov,E;Kopras,E;Zilberman,A;Periasamy,M
通讯作者：
Periasamy,M
影响因子：
56.9
作者：
BRIGGS, MR;KADONAGA, JT;TJIAN, R
通讯作者：
TJIAN, R
影响因子：
10.5
作者：
PASCAL, E;TJIAN, R
通讯作者：
TJIAN, R
DOI：
10.1073/pnas.88.23.10676
发表时间：
1991-12-01
影响因子：
11.1
作者：
BABIJ, P;KELLY, C;PERIASAMY, M
通讯作者：
PERIASAMY, M