Rapamycin Inhibits Expression of Elongation of Very-long-chain Fatty Acids 1 and Synthesis of Docosahexaenoic Acid in Bovine Mammary Epithelial Cells

Mammalian target of rapamycin complex 1 (mTORC1) is a central regulator of cell growth and metabolism and is sufficient to induce specific metabolic processes, including de novo lipid biosynthesis. Elongation of very-long-chain fatty acids 1 (ELOVL1) is a ubiquitously expressed gene and the product of which was thought to be associated with elongation of carbon (C) chain in fatty acids. In the present study, we examined the effects of rapamycin, a specific inhibitor of mTORC1, on ELOVL1 expression and docosahexaenoic acid (DHA, C22:6 n-3) synthesis in bovine mammary epithelial cells (BMECs). We found that rapamycin decreased the relative abundance of ELOVL1 mRNA, ELOVL1 expression and the level of DHA in a time-dependent manner. These data indicate that ELOVL1 expression and DHA synthesis are regulated by mTORC1 in BMECs

Inhibition of Mammalian Target of Rapamycin Complex 1 (mTORC1) Downregulates ELOVL1 Gene Expression and Fatty Acid Synthesis in Goat Fetal Fibroblasts

Elongation of very-long-chain fatty acids 1 (ELOVL1) is a ubiquitously expressed gene that belongs to the ELOVL family and regulates the synthesis of very-long-chain fatty acids (VLCFAs) and sphingolipids, from yeast to mammals. Mammalian target of rapamycin complex 1 (mTORC1) is a central regulator of cell metabolism and is associated with fatty acids synthesis. In this study, we cloned the cDNA that encodes Cashmere goat (Capra hircus) ELOVL1 (GenBank Accession number KF549985) and investigated its expression in 10 tissues. ELOVL1 cDNA was 840 bp, encoding a deduced protein of 279 amino acids, and ELOVL1 mRNA was expressed in a wide range of tissues. Inhibition of mTORC1 by rapamycin decreased ELOVL1 expression and fatty acids synthesis in Cashmere goat fetal fibroblasts. These data show that ELOVL1 expression is regulated by mTORC1 and that mTORC1 has significant function in fatty acids synthesis in Cashmere goat

SQSTM1/p62 interacts with FKBP38 and regulates cell cycle in Cashmere goat foetal fibroblasts

<p>SQSTM1 (sequestosome 1, also known as p62) is a multifunctional scaffold protein implicated in diverse cell physiology processes, such as autophagy, cell signalling, and protein turnover. FKBP38 (FK506-binding protein 38) is a member of FKBPs family and plays a key role in various cellular processes, including signalling transduction, embryonic development and apoptosis. In order to explore the role of SQSTM1/p62 gene in cell-cycle progression and proliferation of goat foetal fibroblast (GFbs), SQSTM1/p62 gene was cloned and characterized. Furthermore, the yeast two-hybrid screening system was used to identify the interaction between SQSTM1/p62 and FKBP38. The results suggested that SQSTM1/p62 directly interacts with FKBP38. Overexpression vector pIRES-EGFP-SQSTM1/p62 and shRNA eukaryotic expression vector pRNAT-U6.1-shSQSTM1/p62 which harboured siRNA targeting the SQSTM1/p62 mRNA were constructed. The overexpression of SQSTM1/p62 gene in GFbs significantly increase the S-phase cells compared with control cells (<i>p </i>< .05). Furthermore, SQSTM1/p62 gene silencing in GFbs leads to a significant decrease of S-phase cells and cell cycle arrest compared with control cells (<i>p </i>< .05). These data indicate that SQSTM1/p62 gene plays an important role in cell-cycle and proliferation of Cashmere GFbs.</p> <p>Abbreviations: Aba: aureobasidin A; EIF1: eukaryotic translation initiation factor 1; FKBP: FK506-binding protein; FKBP38: FK506-binding protein 38; GFbs: goat foetal fibroblast; mTOR: mammalian target of rapamycin; PDLIM7: PDZ and LIM domain 7; SD: standard dropout; SQSTM1/p62: sequestosome 1</p