A novel processing system of sterol regulatory element-binding protein-1c regulated by polyunsaturated fatty acid

• Published in: Journal of biochemistry (Tokyo) 2014-05, Vol.155 (5), p.301-313
• Main Authors: Nakakuki, Masanori, Kawano, Hiroyuki, Notsu, Tatsuto, Imada, Kazunori, Mizuguchi, Kiyoshi, Shimano, Hitoshi
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Carcinoma, Hepatocellular - metabolism; Carcinoma, Hepatocellular - pathology; Cell Line, Tumor; Fatty Acids, Unsaturated - metabolism; Fatty Acids, Unsaturated - pharmacology; HEK293 Cells - drug effects; Humans; Intracellular Signaling Peptides and Proteins - genetics; Intracellular Signaling Peptides and Proteins - metabolism; Liver Neoplasms - metabolism; Liver Neoplasms - pathology; Male; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mice, Inbred C57BL; Molecular Sequence Data; Mutation; Protein Processing, Post-Translational; Rats; Sterol Regulatory Element Binding Protein 1 - genetics; Sterol Regulatory Element Binding Protein 1 - metabolism; Sterol Regulatory Element Binding Protein 2 - metabolism; Sterols - metabolism; Sterols - pharmacology
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/jb/mvu019

The proteolytic cascade is the key step in transactivation of sterol regulatory element-binding proteins (SREBPs), a transcriptional factor of lipid synthesis. Proteolysis of SREBP-2 is strictly regulated by sterols, but that of SREBP-1c was not strongly sterol-regulated, but inhibited by polyunsaturated fatty acids (PUFAs). In this study, the proteolytic processing of SREBP-1 and -2 was examined by transfection studies of cDNA-encoding mutants in which all the known cleavage sites were disrupted. In cultured cells, sterol-regulated SREBP-2 processing was completely eliminated by mutation of cleavage sites. In contrast, the corresponding SREBP-1c mutants as well as wild type exhibited large amounts of cleaved products in the nuclear extracts from culture cells and murine liver in vivo. The nuclear form of the mutant SREBP-1c was induced by delipidated condition and suppressed by eicosapentaenoic acid, an n-3 PUFA, but not by sterols. This novel processing mechanism was affected by neither SREBP cleavage-activating protein (SCAP) nor insulin-induced gene (Insig)-1, unlike SREBP-2, but abolished by a serine protease inhibitor. Through analysis of deletion mutant, a site-2 protease recognition sequence (DRSR) was identified to be involved in this novel processing. These findings suggest that SREBP-1c cleavage could be subjected to a novel PUFA-regulated cleavage system in addition to the sterol-regulatory SCAP/Insig system.