INVIVO FOOTPRINTING OF A MUSCLE SPECIFIC ENHANCER BY LIGATION MEDIATED PCR
INVIVO FOOTPRINTING OF A MUSCLE SPECIFIC ENHANCER BY LIGATION MEDIATED PCR
复制标题
DOI：
10.1126/science.2814500
复制
发表时间：
1989-11-10
期刊：
影响因子：
56.9
通讯作者：
WOLD, B
中科院分区：
文献类型：
--
作者：
MUELLER, PR;WOLD, B
在体内的蛋白质-DNA相互作用的发育调节增强子的小鼠肌肉肌酸激酶（MCK）基因进行了检查，通过一个新开发的聚合酶链反应（PCR）足迹法。这种连接介导的单侧PCR技术允许整个序列梯的指数扩增。在MCK基因活跃转录的终末分化肌细胞中检测到几个足迹。在分化前的肌源性细胞或非肌细胞中均未观察到。两个足迹似乎对应于可以在体外结合肌源性调节因子MyoD 1的位点，而另外两个足迹则代表肌肉特异性使用明显的一般因子。由于MyoD 1是由未分化的成肌细胞合成的，因此这些数据意味着在分化之前，额外的调节机制必须限制该蛋白质与其靶位点之间的相互作用。
In vivo protein-DNA interactions at the developmentally regulated enhancer of the mouse muscle creatine kinase (MCK) gene were examined by a newly developed polymerase chain reaction (PCR) footprinting procedure. This ligation mediated, single-sided PCR technique permits the exponential amplification of an entire sequence ladder. Several footprints were detected in terminally differentiated muscle cells where the MCK gene is actively transcribed. None were observed in myogenic cells prior to differentiation or in nonmuscle cells. Two footprints appear to correspond to sites that can bind the myogenic regulator MyoD1 in vitro, whereas two others represent muscle specific use of apparently general factors. Because MyoD1 is synthesized by undifferentiated myoblasts, these data imply that additional regulatory mechanisms must restrict the interaction between this protein and its target site prior to differentiation.