Chen Wei, Xu Houqiang, Chen Xiang, Liu Zhongwei, Zhang Wen, Xia Dan
Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Guizhou University, Guiyang 550025, China.
College of Animal Science, Guizhou University, Guiyang 550025, China.
Int J Mol Sci. 2016 May 5;17(5):682. doi: 10.3390/ijms17050682.
Uncoupling protein 3 (UCP3) is mainly expressed in muscle. It plays an important role in muscle, but less research on the regulation of cattle UCP3 has been performed. In order to elucidate whether cattle UCP3 can be regulated by muscle-related factors, deletion of cattle UCP3 promoter was amplified and cloned into pGL3-basic, pGL3-promoter and PEGFP-N3 vector, respectively, then transfected into C2C12 myoblasts cells and UCP3 promoter activity was measured using the dual-Luciferase reporter assay system. The results showed that there is some negative-regulatory element from -620 to -433 bp, and there is some positive-regulatory element between -433 and -385 bp. The fragment (1.08 kb) of UCP3 promoter was cotransfected with muscle-related transcription factor myogenic regulatory factors (MRFs) and myocyte-specific enhancer factor 2A (MEF2A). We found that UCP3 promoter could be upregulated by Myf5, Myf6 and MyoD and downregulated by MyoG and MEF2A.
解偶联蛋白3（UCP3）主要在肌肉中表达。它在肌肉中发挥重要作用，但关于牛UCP3调控的研究较少。为了阐明牛UCP3是否受肌肉相关因子调控，分别扩增牛UCP3启动子缺失片段并克隆到pGL3-basic、pGL3-promoter和PEGFP-N3载体中，然后转染到C2C12成肌细胞中，使用双荧光素酶报告基因检测系统测量UCP3启动子活性。结果表明，在-620至-433 bp之间存在一些负调控元件，在-433至-385 bp之间存在一些正调控元件。将UCP3启动子片段（1.08 kb）与肌肉相关转录因子生肌调节因子（MRFs）和肌细胞特异性增强因子2A（MEF2A）共转染。我们发现UCP3启动子可被Myf5、Myf6和MyoD上调，被MyoG和MEF2A下调。
