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首页> 外文期刊>Canadian Journal of Physiology and Pharmacology >The single-strand DNA/RNA-binding protein, Purbeta, regulates serum response factor (SRF)-mediated cardiac muscle gene expression.
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The single-strand DNA/RNA-binding protein, Purbeta, regulates serum response factor (SRF)-mediated cardiac muscle gene expression.

机译：单链DNA / RNA结合蛋白Purbeta调节血清反应因子（SRF）介导的心肌基因表达。

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摘要

Single-strand DNA-binding proteins, Puralpha and Purbeta, play a role in cell growth and differentiation by modulating both transcriptional and translational controls of gene expression. We have previously characterized binding of Puralpha and Purbeta proteins to a purine-rich negative regulatory (PNR) element of the rat cardiac alpha-myosin heavy chain (MHC) gene that controls cardiac muscle specificity. In this study we investigated the role of upstream sequences of the alpha-MHC promoter in Purbeta-mediated gene repression. In the transient transfection analysis overexpression of Purbeta revealed a negative regulatory effect on serum response factor (SRF)-dependent alpha-MHC and alpha-skeletal actin expression in muscle cell background. Contrary, in nonmuscle cells, Purbeta showed no repressive effect. The results obtained from gel-shift assays demonstrated a sequence specific competitive binding of Purbeta to the minus strand of the SRF-binding, CArG box sequences of different muscle genes, but notto the SRF-binding, SRE sequences of the c-fos gene. These element-specific associations of Purbeta with muscle CArG boxes may, in part, explain why muscle gene expression is downregulated in disease states in which Purbeta levels are elevated. This data also provide a mechanistic distinction between muscle CArG boxes and nonmuscle serum response element (SRE) sequences in terms of their affinity to bind to SRF and their ability to regulate cell-specific gene expression.
机译：单链DNA结合蛋白Puralpha和Purbeta通过调节基因表达的转录和翻译控制，在细胞生长和分化中发挥作用。我们以前已经表征了Puralpha和Purbeta蛋白与大鼠心脏α-肌球蛋白重链（MHC）基因的富含嘌呤的负调节（PNR）元件的结合，该基因控制心肌的特异性。在这项研究中，我们调查了α-MHC启动子的上游序列在Purbeta介导的基因阻抑中的作用。在瞬时转染分析中，Purbeta的过表达揭示了对肌肉细胞背景中血清反应因子（SRF）依赖性α-MHC和α-骨骼肌肌动蛋白表达的负调控作用。相反，在非肌肉细胞中，Purbeta没有显示出抑制作用。从凝胶移位分析获得的结果表明，Purbeta与不同肌肉基因的SRF结合CArG盒序列的负链具有序列特异性竞争结合，但与c-fos基因的SRF结合SRE序列没有竞争性结合。 Purbeta与肌肉CArG框的这些元素特定关联可能部分解释了为什么在Purbeta水平升高的疾病状态下，肌肉基因表达被下调。该数据还提供了肌肉CArG盒与非肌肉血清反应元件（SRE）序列之间的机械区别，即它们与SRF结合的亲和力和调节细胞特异性基因表达的能力。

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