The Expression of Porcine Prdx6 Gene Is Up-Regulated by C/EBPβ and CREB

Xinyu Wu; Panlong Ji; Liang Zhang; Guowei Bu; Hao Gu; Xiaojing Wang; Yuanzhu Xiong; Bo Zuo

doi:10.1371/journal.pone.0144851

The Expression of Porcine Prdx6 Gene Is Up-Regulated by C/EBPβ and CREB

PLoS One. 2015 Dec 14;10(12):e0144851. doi: 10.1371/journal.pone.0144851. eCollection 2015.

Authors

Xinyu Wu¹, Panlong Ji¹, Liang Zhang¹, Guowei Bu¹, Hao Gu¹, Xiaojing Wang², Yuanzhu Xiong¹, Bo Zuo¹

Affiliations

¹ Key Laboratory of Swine Genetics and Breeding, Ministry of Agriculture and Key Lab of Agricultural Animal Genetics and Breeding, Ministry of Education, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, P.R. China.
² Department of Basic Veterinary Physiology and Biochemistry Laboratory, Huazhong Agricultural University, Wuhan, 430070, PR China.

Abstract

Peroxiredoxin6 (Prdx6) is one of the peroxiredoxin (Prdxs) family members that play an important role in maintaining cell homeostasis. Our previous studies demonstrated that Prdx6 was significantly associated with pig meat quality, especially meat tenderness. However, the transcriptional regulation of porcine Prdx6 remains unclear. In this study, we determined the transcription start site (TSS) of porcine Prdx6 gene by 5' rapid-amplification of cDNA ends (5' RACE). Several regulatory elements including CCAAT/enhancer-binding proteinβ (C/EBPβ), Myogenic Differentiation (MyoD), cAMP response element binding protein (CREB), stimulating protein1 (Sp1) and heat shock factor (HSF) binding sites were found by computational analyses together with luciferase reporter system. Overexpression and RNA interference experiments showed that C/EBPβ or CREB could up-regulate the expression of porcine Prdx6 gene at both mRNA and protein level. Electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation assays (ChIP) confirmed that C/EBPβ and CREB could interact with Prdx6 promoter. Immuoprecipitation results also showed that C/EBPβ could interact with Prdx6 in vivo. Taken together, our findings identified C/EBPβ and CREB as the important regulators of porcine Prdx6 gene expression, and offered clues for further investigation of Prdx6 gene function.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adipogenesis / genetics
Animals
Base Sequence
Binding Sites
CCAAT-Enhancer-Binding Protein-beta / genetics*
CCAAT-Enhancer-Binding Protein-beta / metabolism
Cell Differentiation
Cell Line
Cyclic AMP Response Element-Binding Protein / genetics*
Cyclic AMP Response Element-Binding Protein / metabolism
Epithelial Cells / cytology
Epithelial Cells / metabolism
Fibroblasts / cytology
Fibroblasts / metabolism
Gene Expression Regulation, Developmental*
Heat-Shock Proteins / genetics
Heat-Shock Proteins / metabolism
Meat / analysis*
Mice
Molecular Sequence Data
Muscle Development / genetics*
MyoD Protein / genetics
MyoD Protein / metabolism
Myoblasts / cytology
Myoblasts / metabolism
Peroxiredoxin VI / genetics*
Peroxiredoxin VI / metabolism
Promoter Regions, Genetic
Protein Binding
Signal Transduction
Sp1 Transcription Factor / genetics
Sp1 Transcription Factor / metabolism
Swine
Transcription Initiation Site

Substances

CCAAT-Enhancer-Binding Protein-beta
Cyclic AMP Response Element-Binding Protein
Heat-Shock Proteins
MyoD Protein
Sp1 Transcription Factor
Peroxiredoxin VI

Grants and funding

This study was financially supported by the Key National High Technology Development Project of China (project no. 2011AA100301), the National Key Technology Support Program of China (grant no. 2014BAD20B01), the Agricultural Innovation Fund of Hubei Province (grant no. 2007-620), the Fundamental Research Funds for the Central Universities (grant no. 2014PY038) and the Key Project of Science and Technology of Zhejiang Province (grant no. 2014C02010).