VPS28通过泛素化信号通路调控奶牛乳腺上皮细胞中乳蛋白的合成

doi:10.11843/j.issn.0366-6964.2022.08.014

Abstract

Abstract: The aim of this study was to analyze the molecular mechanism of VPS28 gene regulating milk protein synthesis and to lay a theoretical foundation for molecular breeding of lactation traits in dairy cows. In this study, the expression level of VPS28 gene in primary mammary epithelial cells (BMECs) of dairy cows was knocked down by RNAi technology, and the expression level of 11 genes, ubiquitin proteins, and the proteasome activity involved in milk protein synthesis, ubiquitin-lysosome, and ubiquitin-proteasome pathways were detected. Then, the activity of proteasome and lysosome in BMECs were inhibited, and the expression levels of casein-related genes and ribosomal proteins were detected. Finally, iTRAQ comparative proteomics was used to analyze the differentially expressed proteins in BMECs before and after knocking down. After VPS28 knockdown, the CSN1S1, CSN2, CSN3, RPS8, UBC, PSMC3, and PSMC5 genes were significantly up-regulated, while PSMD12 was significantly down-regulated. After proteasome inhibition, the CSN1S1, CSN2, and CSN3 genes were significantly up-regulated, while RPL13 was significantly down-regulated. The expression of casein-related genes was not significant after lysosome inhibition. iTRAQ results showed that there were 129 differentially expressed proteins, the down-regulated proteins were mainly enriched in ribosomes, lysosomes, spliceosomes,and other related pathways, while the up-regulated proteins were mainly enriched in the protein processing in the endoplasmic reticulation, vasopressin-regulated water reabsorption process, RNA transport, and other pathways. The results indicate that VPS28 gene can affect milk protein synthesis through ubiquitination signaling pathways in BMECs.

Key words: VPS28, milk protein, ubiquitination, RNAi, iTRAQ

CLC Number:

S823.91

REN Yu, YAN Wenquan, DING Yuezhu, HU Mengxue, ZHOU Jielong, WU Peifu, CHEN Fenfen, LIU Lili. VPS28 Regulates Milk Protein Synthesis in Bovine Mammary Epithelial Cells through Ubiquitination Signaling Pathway[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2558-2567.

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VPS28 Regulates Milk Protein Synthesis in Bovine Mammary Epithelial Cells through Ubiquitination Signaling Pathway

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