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MEF2A 和 MyoG 在牛 LATS2 基因转录调控中的作用。

Roles of MEF2A and MyoG in the transcriptional regulation of bovine LATS2 gene.

机构信息

Institute of Animal Sciences, Ningxia Academy of agricultural and Forestry Sciences, Yinchuan 750021, China.

College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.

出版信息

Res Vet Sci. 2022 Dec 20;152:417-426. doi: 10.1016/j.rvsc.2022.08.030. Epub 2022 Sep 5.

Abstract

As an important downstream effector gene in the hippo signaling pathway, large tumor suppressor gene 2 (LATS2) is involved in cell proliferation and differentiation, organ size and tissue regeneration, and plays an important role in regulating the growth and development of animal muscles. The purpose of this study is to explore the temporal expression of bovine LATS2 gene, and determine the key transcription factors for regulating bovine LATS2 gene. The result showed that bovine LATS2 gene was highly expressed in liver and longissimus dorsi, and was up-regulated in infancy muscle. In addition, it was highly expressed on the 2th day during the differentiation stage of myoblast. The upstream 1.7 Kb sequence of the 5 'translation region of bovine LATS2 gene was cloned, and 7 different deletion fragments were amplified by the upstream primers. These fragments were constructed into double luciferase reporter vectors and transfected into myoblasts and myotubes cells, respectively to detect the core promoter regions. In addition, the key transcription factors of the core promoter sequence of the bovine LATS2 gene were analyzed and predicted by online software. Combining with site-directed mutations, siRNA interference and chromatin immunoprecipitation technology, it was identified that MEF2A and MyoG combined in core promoter region (-248/-56) to regulate the transcription activity of bovine LATS2 gene. The results have laid a theoretical foundation for exploring the molecular regulation mechanism of LATS2 gene in the process of muscle growth.

摘要

作为 Hippo 信号通路的一个重要下游效应基因,大肿瘤抑制基因 2(LATS2)参与细胞增殖和分化、器官大小和组织再生,在调节动物肌肉的生长和发育中发挥重要作用。本研究旨在探讨牛 LATS2 基因的时空表达,确定调节牛 LATS2 基因的关键转录因子。结果表明,牛 LATS2 基因在肝脏和背最长肌中高度表达,并在幼肌中上调。此外,它在成肌细胞分化阶段的第 2 天高度表达。克隆了牛 LATS2 基因 5'翻译区上游 1.7 Kb 序列,并用上游引物扩增了 7 个不同的缺失片段。这些片段分别构建成双荧光素酶报告载体,并转染到成肌细胞和肌管细胞中,以检测核心启动子区域。此外,还通过在线软件分析和预测了牛 LATS2 基因核心启动子序列的关键转录因子。结合定点突变、siRNA 干扰和染色质免疫沉淀技术,鉴定出 MEF2A 和 MyoG 在核心启动子区域(-248/-56)结合,调节牛 LATS2 基因的转录活性。该结果为探索 LATS2 基因在肌肉生长过程中的分子调控机制奠定了理论基础。

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