College of Animal Science and Technology & Gansu Key Laboratory of Herbivorous Animal Biotechnology, Gansu Agricultural University, Lanzhou, P. R. China.
College of Animal Science and Technology, Northwest A&F University, Shaanxi, P. R. China.
Anim Biotechnol. 2022 Aug;33(4):757-764. doi: 10.1080/10495398.2020.1837848. Epub 2020 Oct 30.
Intramuscular fat (IMF) content is a key determinant of beef quality, making it a key topic of research interest. ATP5B serves as the catalytic component of the mitochondrial ATP synthase complex and plays essential roles in controlling fat contents and oxidative metabolism in bovine skeletal muscle. In this study, we determined that bovine ATP5B was highly expressed in longissimus thoracis. To elucidate the molecular mechanisms involved in bovine ATP5B regulation, we cloned and characterized the promoter region of ATP5B. Applying 5'-rapid amplification of cDNA end analysis (RACE), we identified two transcriptional start sites (TSSs) in its promoter region. Using a series of 5'-deletion promoter plasmids in luciferase reporter assay, we found that the proximal minimal promoter of ATP5B was located within the region -539/220 relative to the TSS. Site-directed mutation in combination with chromatin immunoprecipitation (ChIP) assays demonstrated that MyoD and GATA1 binding to the promoter region drives bovine ATP5B transcription. Taken together, these results provide new insight into the regulatory mechanisms of ATP5B transcription in mediating the IMF content of beef.
肌内脂肪（IMF）含量是牛肉质量的关键决定因素，因此成为研究的重点。ATP5B 是线粒体 ATP 合酶复合物的催化亚基，在控制牛骨骼肌脂肪含量和氧化代谢方面发挥着重要作用。本研究发现牛 ATP5B 在胸最长肌中高表达。为了阐明与牛 ATP5B 调控相关的分子机制，我们克隆并鉴定了 ATP5B 的启动子区域。通过 5'-快速扩增 cDNA 末端分析（RACE），我们在其启动子区域鉴定了两个转录起始位点（TSS）。通过一系列 5'-缺失启动子质粒的荧光素酶报告基因检测，我们发现 ATP5B 的近端最小启动子位于相对于 TSS 的-539/220 区域内。定点突变结合染色质免疫沉淀（ChIP）实验表明，MyoD 和 GATA1 结合到启动子区域驱动牛 ATP5B 的转录。综上所述，这些结果为调节肌内脂肪含量的 ATP5B 转录的调控机制提供了新的见解。
