敲降VPS28基因对中国荷斯坦奶牛乳脂合成的调控

doi:10.16288/j.yczz.18-134

遗传 ›› 2018, Vol. 40 ›› Issue (12): 1092-1100.doi: 10.16288/j.yczz.18-134

敲降VPS28基因对中国荷斯坦奶牛乳脂合成的调控

刘莉莉¹,郭爱伟¹,吴培福¹,陈粉粉¹,杨亚晋¹,张勤²()

^1. 西南林业大学生命科学学院,昆明 650224
^2. 中国农业大学动物科技学院,畜禽育种国家工程实验室,北京 1000193

收稿日期:2018-05-15 修回日期:2018-07-31 出版日期:2018-12-20 发布日期:2018-10-30
通讯作者: 张勤 E-mail:qzhang@cau.edu.cn
作者简介:刘莉莉,博士,讲师,研究方向：动物遗传育种。E-mail: liulily0518@163.com
基金资助:
国家自然科学基金项目(3120772);国家转基因育种发展专项(2014ZX0800953B);教育部重点学科生物学一级学科建设项目(51400666);云南省教育厅科学研究基金教师类项目(2018JS333)

Regulation of VPS28 gene knockdown on the milk fat synthesis in Chinese Holstein dairy

Lili Liu¹,Aiwei Guo¹,Peifu Wu¹,Fenfen Chen¹,Yajin Yang¹,Qin Zhang²()

^1. College of Life Science, Southwest Forestry University, Kunming 650224, China
^2. Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture,College of Animal Science and Technology, China Agricultural University, Beijing 100193, China

Received:2018-05-15 Revised:2018-07-31 Online:2018-12-20 Published:2018-10-30
Contact: Zhang Qin E-mail:qzhang@cau.edu.cn
Supported by:
Supported by the National Natural Science Foundation of China(3120772);the National Major Development Program of Transgenic Breeding(2014ZX0800953B);Ministry of education key subject biology level discipline construction project(51400666);Yunnan Provincial Department of Education Fund for Scientific Research(2018JS333)

摘要/Abstract

摘要：

本课题组前期通过GWAS研究,发现VPS28基因在荷斯坦奶牛乳腺组织中特异性高表达,且其5′-UTR的突变位点-58C>T与乳脂性状关联,但其对乳脂性状的调控机理尚未明确。本研究为了明确VPS28基因及其突变位点-58C>T对乳脂的调控机理,首先利用启动子活性分析检测突变位点-58C>T对VPS28基因的影响,发现该突变位点显著降低VPS28基因启动子活性;然后利用RNA干扰技术敲降奶牛原代乳腺上皮细胞中VPS28基因表达量,检测VPS28通路和乳脂合成相关基因mRNA表达量以及细胞中脂肪滴形态,分析结果发现敲降VPS28基因可降低泛素化-溶酶体和泛素化-蛋白酶体通路基因和乳脂合成相关基因的表达量,并提高细胞中甘油三酯的合成,预示VPS28基因可能通过泛素化-溶酶体和泛素化-蛋白酶体途径调控乳脂生成。本研究结果在转录组水平揭示VPS28基因对乳脂合成的调控机制,为奶牛乳脂性状的分子育种研究提供参考依据。

关键词: 中国荷斯坦奶牛, VPS28基因, 单核苷酸多态性, 乳脂合成, 调控

Abstract:

In a previous genome-wide association study on milk production traits in Chinese Holstein population, we discovered VPS28 gene was highly expressed in mammary gland tissue. Further, a -58C>T mutantion at the 5°-UTR of the gene was significantly associated with milk fat content traits. However, its regulation on milk fat content is still uncertain. In this study, we explored the effect of this -58C>T mutation on VPS28, and found that it could significantly reduce promoter activity of VPS28 gene. To identify the potential function of this SNP, we performed RNAi experiments to knockdown VPS28 gene in BMECs and examined the general effects of VPS28 knockdown on BMECs. The results showed that VPS28 knockdown could decrease the mRNA expression of genes in ubiquitination-lysosome and ubiquitination- proteasome pathways, increase the mRNA expression of genes in milk fat synthesis pathway and promote triglyceride synthesis in BMECs. Our data indicate that VPS28 gene could regulate milk fat synthesis pathway; and promote triglyceride synthesis in BMECs. Our data indicate that VPS28 gene could regulate milk fat synthesis through modulating the ubiquitination-lysosome and ubiquitination-proteasome systems. Our results demonstrate the molecular mechanism of VPS28 on regulation of milk synthesis in dairy cattle through the ubiquitination signaling pathway, thereby supporting a relationship between milk fat synthesis and ubiquitination and laying the molecular foundation in breeding of dairy cattle in the future.

Key words: Chinese Holstein dairy, VPS28 gene, single nucleotide polymorphism, milk fat related genes, transcriptional regulation

刘莉莉, 郭爱伟, 吴培福, 陈粉粉, 杨亚晋, 张勤. 敲降VPS28基因对中国荷斯坦奶牛乳脂合成的调控[J]. 遗传, 2018, 40(12): 1092-1100.

Lili Liu, Aiwei Guo, Peifu Wu, Fenfen Chen, Yajin Yang, Qin Zhang. Regulation of VPS28 gene knockdown on the milk fat synthesis in Chinese Holstein dairy[J]. Hereditas(Beijing), 2018, 40(12): 1092-1100.

参考文献

[1]	Walther TC, Farese RV . The life of lipid droplets. Biochim Biophys Acta, 2009,1791(6):459-466.
[2]	Heid HW, Keenan TW . Intracellular origin and secretion of milk fat globules. Eur J Cell Biol, 2005,84(2-3):245-258.
[3]	Bauman DE, Mather IH, Wall RJ, Lock AL . Major advances associated with the biosynthesis of milk. J Dairy Sci, 2006,89(4):1235-1243.
[4]	Bionaz M, Loor JJ . Gene networks driving bovine milk fat synthesis during the lactation cycle. BMC Genomics, 2008,9:366.
[5]	McManaman JL, Russell TD, Schaack J, Orlicky DJ, Robenek H . Molecular determinants of milk lipid secretion. J Mammary Gland Biol Neoplasia, 2007,12(4):259-268.
[6]	Jiang L, Liu X, Yang J, Wang H, Jiang J, Liu L, He S, Ding X, Liu J, Zhang Q . Targeted resequencing of GWAS loci reveals novel genetic variants for milk production traits. BMC Genomics, 2014,15:1105.
[7]	Raiborg C, Stenmark H . The ESCRT machinery in endosomal sorting of ubiquitylated membrane proteins. Nature, 2009,458(7237):445-452.
[8]	Elia N, Sougrat R, Spurlin TA, Hurley JH, Lippincott- Schwartz J . Dynamics of endosomal sorting complex required for transport (ESCRT) machinery during cytokinesis and its role in abscission. Proc Natl Acad Sci USA, 2011,108(12):4846-4851.
[9]	Agromayor M, Soler N, Caballe A, Kueck T, Freund SM, Allen MD, Bycroft M, Perisic O, Ye Y , McDonald B,Scheel H, Hofmann K, Neil SJ, Martin-Serrano J, Williams RL. The UBAP1 subunit of ESCRT-I interacts with ubiquitin via a SOUBA domain. Structure, 2012,20(3):414-428.
[10]	Hoeller D, Dikic I . Targeting the ubiquitin system in cancer therapy. Nature, 2009,458(7237):438-444.
[11]	Winter V, Hauser MT . Exploring the ESCRTing machinery in eukaryotes. Trends Plant Sci, 2006,11(3):115-123.
[12]	Saksena S, Sun J, Chu T, Emr SD . ESCRTing proteins in the endocytic pathway. Trends Biochem Sci, 2007,32(12):561-573.
[13]	Hirsch C, Gauss R, Horn SC, Neuber O, Sommer T . The ubiquitylation machinery of the endoplasmic reticulum. Nature, 2009,458(7237):453-460.
[14]	Katzmann DJ, Babst M, Emr SD . Ubiquitin-dependent sorting into the multivesicular body pathway requires the function of a conserved endosomal protein sorting complex, ESCRT-I. Cell, 2001,106(2):145-155.
[15]	Sevrioukov EA, Moghrabi N, Kuhn M, Krämer H . A mutation in dVps28 reveals a link between a subunit of the endosomal sorting complex required for transport-I complex and the actin cytoskeleton in Drosophila. Mol Biol Cell, 2005,16(5):2301-2312.

[1]	吕倩雯, 杨永芳. 植物小肽信号生物学功能及其在作物改良中研究进展[J]. 遗传, 2023, 45(9): 813-828.
[2]	于一凡, 欧阳臻, 郭娟, 赵瑜君, 黄璐琦. 植物质体基因工程调控元件研究进展[J]. 遗传, 2023, 45(6): 501-513.
[3]	王舜泽, 江丰, 朱东丽, 杨铁林, 郭燕. Hi-C技术在三维基因组学和疾病致病机理研究中的应用[J]. 遗传, 2023, 45(4): 279-294.
[4]	张茜, 王子豪, 田烨. 跨组织线粒体应激信号交流调控机体衰老研究进展[J]. 遗传, 2023, 45(3): 187-197.
[5]	王承贤, 容益康, 崔敏. 果蝇限制端粒转座子的分子机制[J]. 遗传, 2023, 45(3): 221-228.
[6]	吴丹丹, 朱明昆, 方忠艳, 马伟. 植物B染色体的分子结构组成及遗传机制研究进展[J]. 遗传, 2022, 44(9): 772-782.
[7]	徐思远, 寿佳, 吴强. HS5-1增强子eRNA PEARL对原钙粘蛋白α基因簇的表达调控[J]. 遗传, 2022, 44(8): 695-764.
[8]	谢甜, 王梅, 高瑞钰, 苗艳尼, 张燚铭, 蒋婧. 光控诱导重组系统的开发与应用[J]. 遗传, 2022, 44(8): 655-671.
[9]	郝艳, 雷富民. 适应性演化的分子遗传机制：以高海拔适应为例[J]. 遗传, 2022, 44(8): 635-654.
[10]	罗熹晨, 刘慧, 刘学英, 李欣欣, 廖红, 傅向东. 植物响应环境变化的长距离信号传导[J]. 遗传, 2022, 44(7): 556-566.
[11]	赵岩, 王晨鑫, 杨天明, 李春爽, 张丽宏, 杜冬妮, 王若曦, 王静, 魏民, 巴雪青. DNA氧化损伤8-羟鸟嘌呤与肿瘤的发生发展[J]. 遗传, 2022, 44(6): 466-477.
[12]	张元, 赵语婷, 庄乐南, 贺津. 转录中介体复合物在心血管发育和疾病中的转录调控作用[J]. 遗传, 2022, 44(5): 383-397.
[13]	漆思晗, 王棨临, 张俊有, 刘倩, 李春燕. 增强子调控癌症发生发展的机制研究[J]. 遗传, 2022, 44(4): 275-288.
[14]	曲卉, 柳毅, 陈雅文, 汪晖. 环境因素所致印迹基因改变与子代器官发育[J]. 遗传, 2022, 44(2): 107-116.
[15]	刘聪, 冯佳妮, 李玮玮, 朱伟伟, 薛云新, 王岱, 赵西林. 细胞dNTP库的稳态维持与基因组稳定性[J]. 遗传, 2022, 44(2): 96-106.

Metrics

www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:

敲降VPS28基因对中国荷斯坦奶牛乳脂合成的调控

Regulation of VPS28 gene knockdown on the milk fat synthesis in Chinese Holstein dairy

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

编辑推荐

Metrics