Molecular and Cellular Biochemistry

, Volume 363, Issue 1–2, pp 291–299 | Cite as

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

  • Bing Deng
  • Jianghui Wen
  • Yi Ding
  • Qishuang Gao
  • Haijun Huang
  • Zhiping Ran
  • Yunguo Qian
  • Jian Peng
  • Siwen Jiang
Article

Abstract

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 PPARγ but downregulated by C/EBPα and C/EBPβ.

Keywords

MSTN Promoter Transcriptional activity Adipogenesis Myogenesis 

Notes

Acknowledgments

This research was supported by the major projects of Genetically Modified Animals (2009ZX08009-161B), the creative team project of Chinese education ministry (IRT-0831) and National Natural Science Foundation of China (30871779).

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Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Bing Deng
    • 1
    • 2
  • Jianghui Wen
    • 3
  • Yi Ding
    • 1
  • Qishuang Gao
    • 2
  • Haijun Huang
    • 2
  • Zhiping Ran
    • 2
  • Yunguo Qian
    • 2
  • Jian Peng
    • 4
  • Siwen Jiang
    • 1
  1. 1.Agricultural Ministry Key Laboratory of Swine Breeding and Genetics and Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education, College of Animal Science and TechnologyHuazhong Agricultural UniversityWuhanPeople’s Republic of China
  2. 2.Institute of Animal Science and Veterinary Medicine of WuhanWuhan Academy of Agricultural Science & TechnologyWuhanPeople’s Republic of China
  3. 3.School of Mathematics and StatisticsHuazhong University of Science and TechnologyWuhanPeople’s Republic of China
  4. 4.Department of Animal Nutrition and Feed Science, College of Animal Science and TechnologyHuazhong Agricultural UniversityWuhanPeople’s Republic of China

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