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首页> 外文期刊>Molecular and Cellular Biology >A 29-nucleotide DNA segment containing an evolutionarily conserved motif is required in cis for cell-type-restricted repression of the chicken alpha-smooth muscle actin gene core promoter.
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A 29-nucleotide DNA segment containing an evolutionarily conserved motif is required in cis for cell-type-restricted repression of the chicken alpha-smooth muscle actin gene core promoter.

机译：鸡的α-平滑肌肌动蛋白基因核心启动子的细胞类型限制性阻抑，顺式需要一个包含进化保守基序的29个核苷酸的DNA片段。

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A series of 5' deletion mutations of the upstream flanking sequences of the chicken alpha-smooth muscle (aortic) actin gene was prepared and inserted into the chloramphenicol acetyltransferase expression vector pSV0CAT. Deletion recombinants were transfected into fibroblasts, which actively express the alpha-smooth muscle actin gene, and primary myoblast cultures, which accumulate much lower quantities of alpha-smooth muscle actin mRNAs. The first 122 nucleotides of 5'-flanking DNA were found to contain a "core" promoter capable of accurately directing high levels of transcription in both fibroblasts and myotubes. The activity of this core promoter is modulated in fibroblasts by a "governor" element(s) located at least in part between nucleotides -257 and -123. This region contains sequences potentially conserved between mammalian and avian alpha-smooth muscle actin genes as well as one of a pair of 16-base-pair inverted CCAAT box-associated repeats which are conserved among all chordate muscle actin genes examined to date. A smaller DNA segment (-151 to -123) containing these upstream CCAAT box-associated repeats was sufficient to suppress expression of the core promoter in muscle cultures, suggesting that the upstream CCAAT box-associated repeats play a negative role in the alpha-smooth muscle actin gene promoter.
机译：制备了鸡α-平滑肌（主动脉）肌动蛋白基因上游侧翼序列的一系列5'缺失突变，并将其插入到氯霉素乙酰基转移酶表达载体pSV0CAT中。缺失的重组体被转染到成纤维细胞和原代成肌细胞中，成纤维细胞积极表达α-平滑肌肌动蛋白基因，而α-平滑肌肌动蛋白mRNA的表达量要低得多。发现5'侧翼DNA的前122个核苷酸含有“核心”启动子，该启动子能够精确地指导成纤维细胞和肌管中的高水平转录。该核心启动子的活性在成纤维细胞中由至少部分位于核苷酸-257和-123之间的“调控子”元件调节。该区域包含哺乳动物和禽类α-平滑肌肌动蛋白基因之间潜在保守的序列，以及迄今检查的所有胆酸盐肌肌动蛋白基因之间保守的一对16碱基对反向CCAAT盒相关重复序列之一。包含这些上游CCAAT框相关重复序列的较小DNA片段（-151至-123）足以抑制核心培养物在肌肉培养物中的表达，这表明上游CCAAT框相关重复序列在α平滑蛋白中起负作用肌动蛋白基因启动子。

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