YAP 1基因调控新西兰白兔肌肉生长发育  

YAP 1 Regulating the Muscle Growth and Development in New Zealand White Rabbits

作　　者：刘雨晴 陈孟娟 王亿杰 许会芬 李明[1] 于光晴 LIU Yu-Qing;CHEN Meng-Juan;WANG Yi-Jie;XU Hui-Fen;LI Ming;YU Guang-Qing(College of Animal Science and Technology,Henan Agricultural University,Zhengzhou 450000,Henan,China)

机构地区：[1]河南农业大学动物科技学院,郑州450000

出　　处：《中国生物化学与分子生物学报》2023年第12期1765-1775,共11页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然基金青年项目(No.32102833);中国博士后科学基金第70批面上项目(No.2021M701107);河南农业大学大学生创新创业项目(No.2022DC051)资助

摘　　要：Yes相关蛋白1(Yes-associated protein 1,YAP1)在哺乳动物组织中对于调节器官大小、肿瘤发生和细胞的增殖、分化、凋亡等具有重要作用。为探究YAP 1基因在新西兰白兔肌肉组织生长发育中的生物学功能,本研究通过RT-PCR从新西兰白兔腿肌中克隆到YAP 1基因,并对其生物信息学功能、表达模式及亚细胞定位、启动下游基因表达的转录活性,以及参与调控肌肉组织生长发育的功能进行分析。结果表明,YAP1的CDS区序列长度为1302 bp,编码433个氨基酸,等电点为5.10的亲水性蛋白质,YAP1蛋白不存在信号肽和跨膜区域,其结构主要由无规则卷曲构成。系统进化树分析表明,与绵羊的亲缘关系较近,与小鼠的亲缘关系较远。亚细胞定位显示,该蛋白质在细胞核和细胞质中均有表达。RT-qPCR结果显示,YAP 1基因在新西兰白兔的心、肝、脾、肺、肾、卵巢、子宫和肌肉中均有表达,但在肌肉中的表达量最高,且在腿肌组织中表达水平较高。双荧光素酶报告基因检测结果表明,YAP 1基因第127位的丝氨酸突变为丙氨酸(YAP1-S127A),8xGTIIC的启动子活性明显增强。在肌肉细胞中过表达YAP 1基因可以加快肌肉细胞的增殖,同时上调肌球蛋白重链6(myosin heavy chain 6,MYH6)和肌细胞生成素(myogenin,MYOG)基因的mRNA表达水平,以及MYH6和肌分化因子(myogenic differentiation,MYOD)蛋白质水平,抑制肌肉生长抑制素(myostatin,MSTN)基因mRNA的表达水平。综上结果提示,YAP 1基因在新西兰白兔肌肉生长发育中发挥重要作用。本研究为后续进一步阐明YAP 1基因精准调控肌肉生长发育的分子机制奠定基础,同时为改良家兔品种提供理论依据和基因资源。Yes associated protein 1(YAP1)plays an important role in regulating the organ size,tumor development,cell proliferation,differentiation,and apoptosis in mammalian tissues.In this study,in order to explore the biological function of YAP1 for muscle growth and development in New Zealand white rabbit.The YAP1 gene was cloned from the leg muscle of the New Zealand white rabbit by RT-PCR,and we analyzed its function by bioinformatics,its expression pattern and subcellular localization,its transcriptional activity and functions involved in regulating muscle tissue growth and development.The results showed that the length of YAP1 was 1302 bp,encoding 433 amino acids.It is a hydrophilic protein with an Isoelectric point of 5.10.There was no signal peptide or transmembrane region in YAP1,and its structure was mainly composed of irregular curls.The phylogenetic tree analysis showed that it was closely related to sheep and distantly related to mice.Subcellular localization analysis revealed that it is expressed in both the nucleus and cytoplasm.RT-qPCR results indicated that YAP1 was expressed in the heart,liver,spleen,lungs,kidneys,ovaries,uterus,and muscles of New Zealand white rabbits,but the expression level was the highest in leg muscle tissues.Dual luciferase assays showed that the promoter activity of 8xGTIIC was significantly enhanced by mutating Ser-127 to Ala(YAP1-S127A).Overexpression of YAP1 in muscle cells can accelerate the proliferation of muscle cells,while upregulating the mRNA expression levels of MYH6 and MYOG genes,and MYH6 and MYOD protein levels,and inhibiting the mRNA expression level of MSTN.The above results suggest that YAP1 plays an important role in the muscle growth and development of New Zealand white rabbits.This study lays the foundation for further elucidating the molecular mechanism by which YAP1 regulates muscle growth and development,and providing a theoretical basis and genetic resources for improving rabbit breeds.

关 键 词：Yes相关蛋白1 肌肉 家兔 亚细胞定位 转录活性 

分 类 号：Q78[生物学—分子生物学]