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首页> 外文期刊>Molecular and Cellular Biology >E-box sites and a proximal regulatory region of the muscle creatine kinase gene differentially regulate expression in diverse skeletal muscles and cardiac muscle of transgenic mice.
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E-box sites and a proximal regulatory region of the muscle creatine kinase gene differentially regulate expression in diverse skeletal muscles and cardiac muscle of transgenic mice.

机译：E-box位点和肌肉肌酸激酶基因的近端调节区差异性调节转基因小鼠骨骼肌和心肌的表达。

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Previous analysis of the muscle creatine kinase (MCK) gene indicated that control elements required for transcription in adult mouse muscle differed from those required in cell culture, suggesting that distinct modes of muscle gene regulation occur in vivo. To examine this further, we measured the activity of MCK transgenes containing E-box and promoter deletions in a variety of striated muscles. Simultaneous mutation of three E boxes in the 1,256-bp MCK 5' region, which abolished transcription in muscle cultures, had strikingly different effects in mice. The mutations abolished transgene expression in cardiac and tongue muscle and caused a reduction in expression in the soleus muscle (a muscle with many slow fibers) but did not affect expression in predominantly fast muscles: quadriceps, abdominals, and extensor digitorum longus. Other regulatory sequences with muscle-type-specific activities were found within the 358-bp 5'-flanking region. This proximal region conferred relatively strong expression in limb and abdominal skeletal muscles but was inactive in cardiac and tongue muscles. However, when the 206-bp 5' enhancer was ligated to the 358-bp region, high levels of tissue-specific expression were restored in all muscle types. These results indicate that E boxes and a proximal regulatory region are differentially required for maximal MCK transgene expression in different striated muscles. The overall results also imply that within skeletal muscles, the steady-state expression of the MCK gene and possibly other muscle genes depends on transcriptional mechanisms that differ between fast and slow fibers as well as between the anatomical and physiological attributes of each specific muscle.
机译：先前对肌肉肌酸激酶（MCK）基因的分析表明，成年小鼠肌肉中转录所需的控制元件与细胞培养中所需的控制元件不同，这表明在体内发生了不同的肌肉基因调控模式。为了进一步检查这一点，我们在各种横纹肌中测量了包含E-box和启动子缺失的MCK转基因的活性。在1,256-bp MCK 5'区域中三个E盒的同时突变，消除了肌肉培养物中的转录，对小鼠产生了截然不同的作用。该突变消除了心脏和舌头肌肉中的转基因表达，并导致比目鱼肌（一种具有许多慢纤维的肌肉）中的表达减少，但并未影响主要为快肌（股四头肌，腹部和趾长伸肌）的表达。在358 bp的5'侧翼区域内发现了具有肌肉类型特异性活性的其他调控序列。该近端区域在肢体和腹部骨骼肌中表达相对较强，但在心脏和舌头肌肉中不活跃。但是，将206-bp的5'增强子连接到358-bp区域时，在所有肌肉类型中都恢复了高水平的组织特异性表达。这些结果表明，在不同的横纹肌肉中最大的MCK转基因表达需要E盒和近端调节区。总体结果还暗示，在骨骼肌中，MCK基因以及可能其他肌肉基因的稳态表达取决于转录机制，该机制在快纤维和慢纤维之间以及每条特定肌肉的解剖和生理属性之间有所不同。

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