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首页> 外文期刊>Molecular and Cellular Biology >Transcriptional regulation of the muscle creatine kinase gene and regulated expression in transfected mouse myoblasts.
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Transcriptional regulation of the muscle creatine kinase gene and regulated expression in transfected mouse myoblasts.

机译：肌肉肌酸激酶基因的转录调控和转染小鼠成肌细胞中的表达调控。

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The muscle-specific form of creatine kinase (MCK) is induced in differentiating myoblast cultures, and a dramatic increase in mRNA levels precedes and parallels the increase in MCK protein. To study this induction, the complete MCK gene was cloned and characterized. The transcription unit was shown to span 11 kilobases and to contain seven introns. The splice junctions were identified and shown to conform to the appropriate consensus sequences. Close homology with branchpoint consensuses was found upstream of the 3' splice sites in six of seven cases. Transcriptional regulation of the gene in differentiating myoblast cultures was demonstrated by nuclear run-on experiments; increases in transcription accounted for a major part of the increased mRNA levels. Regulated expression of a transfected MCK gene containing the entire transcription unit with 3.3 kilobases of 5'-flanking sequence was also demonstrated during differentiation of the MM14 mouse myoblast cell line. The MCK 5'-flanking region was sufficient to confer transcriptional regulation to a heterologous structural gene, since chloramphenicol acetyl transferase activity was induced during differentiation of cultures transfected with an MCK-chloramphenicol acetyl transferase fusion construct. Examination of the DNA sequence immediately upstream of the transcription start site revealed a 17-nucleotide element which occurred three times. Comparisons with other muscle-specific genes which are also transcriptionally regulated during myogenesis revealed upstream homologies in the alpha-actin and myosin heavy chain genes, but not in the myosin light-chain genes, with the regions containing these repeats. We suggest that coordinate control of a subset of muscle genes may occur via recognition of these common sequences.
机译：肌酸激酶（MCK）的肌肉特异性形式在分化成肌细胞培养物中被诱导，并且mRNA水平的显着增加先于并平行于MCK蛋白的增加。为了研究该诱导，克隆并表征了完整的MCK基因。转录单位显示跨越11千个碱基，并包含七个内含子。剪接连接被鉴定并显示符合适当的共有序列。在7例中的6例中，在3'剪接位点的上游发现了与分支点共有的紧密同源性。通过成核实验证明了该基因在分化成肌细胞培养物中的转录调控。转录的增加占增加的mRNA水平的主要部分。在MM14小鼠成肌细胞系分化过程中，还证实了转染的MCK基因的调控表达，该基因包含具有3.3 kb 5'侧翼序列的整个转录单位。由于在用MCK-氯霉素乙酰转移酶融合构建体转染的培养物的分化过程中诱导了氯霉素乙酰转移酶活性，因此MCK 5'侧翼区足以赋予异源结构基因转录调控。对紧邻转录起始位点上游的DNA序列的检查显示了17个核苷酸的元件，该元件发生了3次。与在成肌过程中也受到转录调控的其他肌肉特异性基因的比较揭示了α-肌动蛋白和肌球蛋白重链基因中上游同源性，而肌球蛋白轻链基因中上游同源性不高，区域包含这些重复序列。我们建议通过识别这些共同序列可能发生的肌肉基因子集的协调控制。

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