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首页> 外文期刊>Molecular and Cellular Biology >A conserved 28-base-pair element (HF-1) in the rat cardiac myosin light-chain-2 gene confers cardiac-specific and alpha-adrenergic-inducible expression in cultured neonatal rat myocardial cells.
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A conserved 28-base-pair element (HF-1) in the rat cardiac myosin light-chain-2 gene confers cardiac-specific and alpha-adrenergic-inducible expression in cultured neonatal rat myocardial cells.

机译：大鼠心肌肌球蛋白轻链2基因中保守的28个碱基对元件（HF-1）在培养的新生大鼠心肌细胞中赋予心脏特异性和α-肾上腺素可诱导的表达。

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To study the transcriptional regulatory mechanisms which mediate cardiac-specific and inducible expression during myocardial cell hypertrophy, we have extensively characterized the rat cardiac myosin light-chain-2 (MLC-2) gene as a model system. The MLC-2 gene encodes a relatively abundant contractile protein in slow skeletal and cardiac muscle and is upregulated during in vivo cardiac hypertrophy and alpha-adrenergic-mediated hypertrophy of neonatal rat myocardial cells. In transient expression assays employing a series of MLC-2-luciferase constructs, recent studies have identified a 250-bp fragment which is sufficient for both cardiac-specific and alpha-adrenergic-inducible expression. Within this 250-bp fragment lie three regions (HF-1, HF-2, and HF-3), each greater than 10 bp in length, which are conserved between the chicken and rat cardiac MLC-2 genes, suggesting their potential role in the regulated expression of this contractile protein gene. As assessed by substitution mutations within each of the conserved regions, the present study demonstrates that HF-1 and HF-2 are important in both cardiac-specific and inducible expression, while HF-3 has no detectable role in the regulated expression of the MLC-2 gene in transient expression assays. HF-1 sequences confer both cardiac-specific and inducible expression to a neutral promoter-luciferase construct but have no significant effect in the skeletal muscle or nonmuscle cell contexts. Thus, these studies have identified a new cardiac-specific regulatory element (HF-1) which plays a role in both cardiac-specific and inducible expression during myocardial cell hypertrophy.
机译：为了研究在心肌细胞肥大过程中介导心脏特异性和诱导型表达的转录调控机制，我们已广泛表征了大鼠心脏肌球蛋白轻链2（MLC-2）基因作为模型系统。 MLC-2基因在慢速骨骼肌和心肌中编码相对丰富的收缩蛋白，在体内心肌肥大和新生大鼠心肌细胞的α-肾上腺素介导的肥大过程中被上调。在采用一系列MLC-2-荧光素酶构建体的瞬时表达测定中，最近的研究已经鉴定出一个250 bp的片段，足以用于心脏特异性和α-肾上腺素诱导的表达。在这个250 bp的片段中，存在三个区域（HF-1，HF-2和HF-3），每个区域的长度都大于10 bp，在鸡和大鼠心脏MLC-2基因之间是保守的，表明它们的潜在作用收缩蛋白基因的调控表达。根据每个保守区内的取代突变评估，本研究表明HF-1和HF-2在心脏特异性表达和诱导型表达中均重要，而HF-3在MLC的调控表达中没有可检测的作用-2基因在瞬时表达分析中。 HF-1序列将心脏特异性表达和诱导型表达赋予中性启动子-荧光素酶构建体，但在骨骼肌或非肌肉细胞环境中没有显着影响。因此，这些研究已经确定了一种新的心脏特异性调控元件（HF-1），它在心肌细胞肥大过程中在心脏特异性表达和诱导型表达中均发挥作用。

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