Vaspin is a novel target gene of hepatic CCAAT-enhancer-binding protein

Aibara, Daisuke; Matsuo, Kohei; Yamano, Shigeru; Matsusue, Kimihiko

doi:10.1016/j.gene.2019.144113

Cited by 12 publications

(12 citation statements)

References 19 publications

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“…The RT-qPCR analysis was performed using liverspecific C/EBPα knockout mice in the C57BL/6J (OB/OB-C/EBPαLKO) or ob/ob C57BL/6J (ob/ob-C/ EBPαLKO) backgrounds to determine the contribution of hepatic C/EBPα to Lipn expression in the mouse liver. A previous study demonstrated that the c/ebpa mRNA level in the OB/OB-or ob/ob-C/EBPαLKO mouse liver was very low at 12% or 5% compared with that in the OB/OB-or ob/ob-C/EBPαWT mouse liver, respectively [11]. The expression of Lipn mRNA was undetectable in the liver of OB/OB-C/EBPαWT mice but was considerably induced in the fatty liver of ob/ob-C/EBPαWT mice (Fig.…”

Section: C/ebpα Regulates the Expression Of Lipn Gene In The Mouse Livermentioning

confidence: 96%

“…3A). CEBPRE-1, CEBPRE-2, CEBPRE-3, and CEBPRE-4 showed 100%, 89%, 78%, and 89% homology with the typical CEBPRE consensus sequence [11], respectively. Lipn luciferase reporter plasmids (-2,967/+27) containing CEBPRE1-4 were transfected into HEK293FT cells with or without C/EBPα or C/EBPβ expression plasmids (Fig.…”

Section: Four Putative Cebpres In the 5'-upstream Region Of Lipnmentioning

confidence: 99%

“…The reverse primer sequence, which was common for all constructs, was as follows: 5'-GGCTGCAGTGACAGTTAGAAGC-3'. Internal deletion constructs for the Lipn 5'-upstream region were prepared by inverse PCR as previously described [10,11]. The Lipn D3 construct was used as a template.…”

Section: Construction Of Reporter and Expression Plasmidsmentioning

confidence: 99%

“…The gels were exposed to an imaging plate (Fujifilm, Tokyo, Japan) and visualized using an FLA-7000 imaging analyzer (Fujifilm). Probe sequences for the EMSA assay were as follows: Vaspin CEBPRE as the control probe [11], 5'-A GAAAGAAAGTTGCAAAAGGTATG-3'; CEBPRE-3, 5'-CCACTATTTCCCAACCTACAAGTG-3'; CEBPRE-4, 5'-GTGTCATTTTCCAAGGTTTCCTCA-3'.…”

Section: Electrophoretic Mobility Shift Assaymentioning

confidence: 99%

“…To examine whether C/EBPα/β directly binds to CEBPRE-3 and CEBPRE-4, EMSA using CEBPRE-3/-4 probes was performed. C/EBPα directly binds to CEBPRE of the vaspin gene as a positive control (control-CEBPRE) [11] and CEBPRE-3. The binding of C/EBPα to CEBPRE-3 was supershifted by anti-C/EBPα IgG.…”

Section: C/ebpα/β Directly Binds To Cebpre-3 In the Lipn Promoter Regionmentioning

confidence: 99%

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Lipase family member N is a novel target gene for CCAAT/enhancer-binding protein α in type 2 diabetic model mouse liver

2022

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CCAAT/enhancer-binding protein α (C/EBPα) is a transcription factor abundantly expressed in the liver and white adipose tissue (WAT). In this study, we investigated the mechanism by which C/EBPα regulates the lipase family member N (Lipn) gene in the mouse liver. Mouse Lipn consists of non-coding exon 1 and the translation start site located in exon 2. Lipn expression in the fatty liver of ob/ob mice was significantly higher than that in OB/OB mice and was significantly repressed by liver-specific C/EBPα deficiency. Lipn expression in ob/ob mice was detected in the liver, epididymal WAT (eWAT), subcutaneous WAT (sWAT), brown adipose tissue (BAT), and skeletal muscle, but not in the kidney, brain, and heart. Lipn expression in the liver, eWAT, and sWAT of wild-type mice was undetectable, although C/EBPα was highly expressed in these tissues. The database analysis revealed four putative C/EBP-responsive elements (CEBPREs), highly homologous with the typical CEBPRE consensus sequence at positions -2,686/-2,678, -1,364/-1,356, -106/-98, and -45/-37 from the transcription start site (+1) of Lipn. Reporter assays using reporter constructs with serial or internal deletions of the 5'-flanking regions of Lipn showed that two functional CEBPREs (-106/-98 and -45/-37) in the Lipn promoter region are essential for enhancing Lipn transcriptional activity by C/EBPα. Electrophoretic mobility shift assay showed that C/EBPα/β binds to CEBPRE (-106/-98). These results suggest that C/EBPα and type 2 diabetic environment may be required for hepatic Lipn expression.

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Section: C/ebpα Regulates the Expression Of Lipn Gene In The Mouse Livermentioning

confidence: 96%

Section: Four Putative Cebpres In the 5'-upstream Region Of Lipnmentioning

confidence: 99%

Section: Construction Of Reporter and Expression Plasmidsmentioning

confidence: 99%

Section: Electrophoretic Mobility Shift Assaymentioning

confidence: 99%

Section: C/ebpα/β Directly Binds To Cebpre-3 In the Lipn Promoter Regionmentioning

confidence: 99%

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