Chen Zongchang, Li Jingsheng, Bai Yanbin, Liu Zhanxin, Wei Yali, Guo Dashan, Jia Xue, Shi Bingang, Zhang Xiaolan, Zhao Zhidong, Hu Jiang, Han Xiangmin, Wang Jiqing, Liu Xiu, Li Shaobin, Zhao Fangfang
Gansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China.
Int J Mol Sci. 2024 Feb 21;25(5):2506. doi: 10.3390/ijms25052506.
Muscle formation directly determines meat production and quality. The non-SMC condensin I complex subunit G (NCAPG) is strongly linked to the growth features of domestic animals because it is essential in controlling muscle growth and development. This study aims to elucidate the tissue expression level of the bovine gene, and determine the key transcription factors for regulating the bovine gene. In this study, we observed that the bovine gene exhibited high expression levels in longissimus dorsi and spleen tissues. Subsequently, we cloned and characterized the promoter region of the bovine gene, consisting of a 2039 bp sequence, through constructing the deletion fragment double-luciferase reporter vector and site-directed mutation-identifying core promoter region with its key transcription factor binding site. In addition, the key transcription factors of the core promoter sequence of the bovine gene were analyzed and predicted using online software. Furthermore, by integrating overexpression experiments and the electrophoretic mobility shift assay (EMSA), we have shown that cAMP response element binding protein 1 (CREB1) and myogenic differentiation 1 (MYOD1) bind to the core promoter region (-598/+87), activating transcription activity in the bovine gene. In conclusion, these findings shed important light on the regulatory network mechanism that underlies the expression of the gene throughout the development of the muscles in beef cattle.
肌肉形成直接决定肉的产量和品质。非SMC凝聚素I复合体亚基G(NCAPG)与家畜的生长特性密切相关,因为它在控制肌肉生长发育中至关重要。本研究旨在阐明牛基因的组织表达水平,并确定调控牛基因的关键转录因子。在本研究中,我们观察到牛基因在背最长肌和脾脏组织中呈现高表达水平。随后,我们通过构建缺失片段双荧光素酶报告载体并利用定点突变鉴定核心启动子区域及其关键转录因子结合位点,克隆并鉴定了牛基因的启动子区域,其由一段2039 bp的序列组成。此外,利用在线软件对牛基因核心启动子序列的关键转录因子进行了分析和预测。进一步地,通过整合过表达实验和电泳迁移率变动分析(EMSA),我们表明环磷酸腺苷反应元件结合蛋白1(CREB1)和成肌分化1(MYOD1)与核心启动子区域(-598/+87)结合,激活牛基因的转录活性。总之,这些发现为肉牛肌肉发育过程中该基因表达的调控网络机制提供了重要线索。