Functional analysis of pig myostatin gene promoter with some adipogenesis- and myogenesis-related factors

• Published in: Molecular and cellular biochemistry 2012-04, Vol.363 (1-2), p.291-299
• Main Authors: Deng, Bing, Wen, Jianghui, Ding, Yi, Gao, Qishuang, Huang, Haijun, Ran, Zhiping, Qian, Yunguo, Peng, Jian, Jiang, Siwen
• Format: Article
• Language: English
• Subjects: 1-Methyl-3-isobutylxanthine - pharmacology; adipogenesis; Adipogenesis - drug effects; Adipogenesis - genetics; Animals; Binding sites; Biochemistry; Biomedical and Life Sciences; Cardiology; CCAAT-Enhancer-Binding Protein-alpha - genetics; CCAAT-Enhancer-Binding Protein-alpha - metabolism; CCAAT-Enhancer-Binding Protein-beta - genetics; CCAAT-Enhancer-Binding Protein-beta - metabolism; CCAAT/enhancer-binding protein; Cell Line; Cloning, Molecular; Dexamethasone; Dexamethasone - pharmacology; Differentiation; Dose-Response Relationship, Drug; Expression vectors; Gene Expression Regulation; Genes; Genes, Reporter; Genetic research; Hogs; Insulin; Insulin - pharmacology; Life Sciences; Luciferase; Medical Biochemistry; Mesenchymal Stromal Cells - metabolism; Mice; Multipotent Stem Cells - metabolism; Muscle Development - genetics; Muscles; Muscular system; Myoblasts - metabolism; MyoD protein; MyoD Protein - genetics; MyoD Protein - metabolism; myostatin; Myostatin - genetics; Myostatin - metabolism; Nucleotides; Oncology; Peroxisome proliferator-activated receptors; PPAR gamma - agonists; PPAR gamma - genetics; Promoter Regions, Genetic - drug effects; Promoters; Protein binding; rosiglitazone; Swine; Thiazolidinediones - pharmacology; Transcription factors; Transcriptional Activation; Transfection
• ISSN: 0300-8177; 1573-4919
• DOI: 10.1007/s11010-011-1181-y

Myostatin (MSTN) is primarily expressed in muscle and plays an important role in muscle and fat development in pigs. However, there is little information about the regulation of pig MSTN . In order to elucidate whether pig MSTN could be regulated by muscle- and fat-related factors, the porcine MSTN promoter was amplified and cloned into pGL3-basic vector, and transfected into cells to analyze the transcriptional activity of promoter with muscle- and fat-related factors through Dual-luciferase reporter assays. 5′-deletion expression showed that there was a negative-regulatory region located between nucleotides −1519 and −1236 bp, and there were some positive-regulatory regions located between −1236 and −568 bp. The longest fragment (1.7 kb) was cotransfected with muscle-related transcription factor myogenic differentiation 1 ( MyoD ), resulting in promoter transcriptional activity upregulation. The fragment was treated by the adipogenic agents (DIM) including dexamethasone, insulin, and isobutyl-1-methylxanthine (IBMX). We found that MSTN promoter transcriptional activity can be regulated by IBMX, but not by DIM. CCAAT/enhancer binding protein (C/EBP) α and C/EBPβ, two proteins which are induced by DIM during adipogenesis were cotransfected with the 1.7-kb fragment, respectively, resulting in promoter transcriptional activity downregulation. Treating the fragment with rosiglitazone which induce the expression of peroxisome proliferator-activated receptor γ ( PPARγ ), resulting in promoter transcriptional activity upregulation. Cotransfection experiments confirmed this result. Taken together, we showed that porcine MSTN could be upregulated by IBMX, MyoD, and PPAR γ but downregulated by C/EBPα and C/EBPβ.