Liu Li Li, Guo Ai Wei, Wu Pei Fu, Chen Fen Fen, Yang Ya Jin, Zhang Qin
College of Life Science, Southwest Forestry University, Kunming 650224, China.
Key Laboratory of Animal Genetics and Breeding of Ministry of Agriculture,College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.
Yi Chuan. 2018 Dec 20;40(12):1092-1100. doi: 10.16288/j.yczz.18-134.
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.
在之前一项针对中国荷斯坦牛群体产奶性状的全基因组关联研究中，我们发现VPS28基因在乳腺组织中高表达。此外，该基因5'-UTR处的一个-58C>T突变与乳脂肪含量性状显著相关。然而，其对乳脂肪含量的调控仍不明确。在本研究中，我们探究了这个-58C>T突变对VPS28的影响，发现它能显著降低VPS28基因的启动子活性。为了鉴定这个单核苷酸多态性（SNP）的潜在功能，我们进行了RNA干扰实验以敲低乳腺上皮细胞（BMECs）中的VPS28基因，并检测VPS28基因敲低对BMECs的总体影响。结果表明，VPS28基因敲低可降低泛素化-溶酶体途径和泛素化-蛋白酶体途径中基因的mRNA表达，增加乳脂肪合成途径中基因的mRNA表达，并促进BMECs中的甘油三酯合成。我们的数据表明，VPS28基因可调节乳脂肪合成途径，并促进BMECs中的甘油三酯合成。我们的数据表明，VPS28基因可通过调节泛素化-溶酶体系统和泛素化-蛋白酶体系统来调节乳脂肪合成。我们的结果揭示了VPS28通过泛素化信号通路调控奶牛乳合成的分子机制，从而支持了乳脂肪合成与泛素化之间的关系，并为未来奶牛育种奠定了分子基础。
