A Novel Smooth Muscle-specific Enhancer Regulates Transcription of the Smooth Muscle Myosin Heavy Chain Gene in Vascular Smooth Muscle Cells (*)

A Novel Smooth Muscle-specific Enhancer Regulates Transcription of the Smooth Muscle Myosin Heavy Chain Gene in Vascular Smooth Muscle Cells (*)
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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
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发表时间:
1995
期刊:
The Journal of Biological Chemistry
影响因子:
--
通讯作者:
P. Babij
P. Babij
中科院分区:
--
文献类型:
--
作者:
R. Kallmeier;C. Somasundaram;P. Babij

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在兔血管平滑肌细胞 (VSMC) 的原代培养物中对兔平滑肌肌球蛋白重链 (SMHC) 基因启动子的 5' 末端缺失突变体进行瞬时 DNA 转染分析。位于转录起始位点上游-1,332位置的阳性元件始终具有最高的相对氯霉素乙酰转移酶(CAT)活性(是最小SMHC启动子的6.3±1.5倍),表明在-1,332和-1,225之间包含额外的107碱基对(bp)DNA片段可以显着增强VSMC中的CAT活性。将突变体转染到几种肌肉和非肌肉细胞系中,没有显示出任何高于对照的显着 CAT 活性,这表明 SMHC 表达需要平滑肌细胞特有的因子。凝胶位移分析表明多种因素与 107-bp 元件相互作用,其中两个因素似乎表现出平滑肌特异性。转染的 VSMC 中增强子功能的测试表明,107 bp 片段表现为经典增强子,即与位置和方向无关。这些结果表明,一种新的 DNA 元件可能调节 SMHC 基因的组织限制性表达,并提供了平滑肌特异性增强子在 VSMC 中作用的第一个例子。
Transient DNA transfection analysis of 5′ end deletion mutants of the rabbit smooth muscle myosin heavy chain (SMHC) gene promoter was performed in primary cultures of rabbit vascular smooth muscle cells (VSMC). A positive element located at position −1,332 upstream of the transcription start site consistently gave the highest relative chloramphenicol acetyltransferase (CAT) activity (6.3 ± 1.5-fold over the minimal SMHC promoter), suggesting that inclusion of the extra 107-base pair (bp) DNA fragment between −1,332 and −1,225 could significantly enhance CAT activity in VSMC. Transfection of mutants into several muscle and nonmuscle cell lines did not show any significant CAT activity above control, showing that factors unique to smooth muscle cells were required for SMHC expression. Gel shift analysis indicated that multiple factors interacted with the 107-bp element, two of which appeared to show smooth muscle specificity. Tests of enhancer function in transfected VSMC indicated that the 107-bp fragment behaved as a classical enhancer, i.e. independently of position and orientation. These results indicate that a novel DNA element may regulate the tissue-restricted expression of the SMHC gene and provides the first example of a role for a smooth muscle-specific enhancer in VSMC.
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: --
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正向和负向顺式作用调节元件介导小鼠血管平滑肌α-肌动蛋白基因的表达。
DOI: --
发表时间: 1992
期刊: The Journal of biological chemistry
影响因子: --
作者:
Foster,DN;Min,B;Foster,LK;Stoflet,ES;Sun,S;Getz,MJ;Strauch,AR
通讯作者: Strauch,AR
DOI: --
发表时间: 1992
期刊: The Journal of biological chemistry
影响因子: --
作者:
Blank,RS;McQuinn,TC;Yin,KC;Thompson,MM;Takeyasu,K;Schwartz,RJ;Owens,GK
通讯作者: Owens,GK