title: The transcription factor ccaat/enhancer binding protein β (C/EBPβ) and miR-27a regulate the expression of porcine Dickkopf2 (DKK2).
abstract: Using Affymetrix porcine Gene-Chip analyses, we found that Dickkopf2 (DKK2), a WNT antagonist, is differentially expressed in pre-ovulatory follicles between Large White and Chinese Taihu sows. This study aims to identify the regulatory factors responsible for DKK2 expression. Deletion fragment and mutation analyses identified DKK2-D3 as the porcine DKK2 core promoter. There were four C/EBPβ binding sites within the DKK2 core promoter. The C allele that results from a spontaneous alteration (DKK2 c.-1130 T > C) in the core promoter was associated with a higher total number born (TNB) and a higher number born alive (NBA) in all parities in a synthetic pig population. This was possibly the result of a change in C/EBPβ binding ability, which was confirmed using chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assays (EMSA). Moreover, C/EBPβ specifically bound to and activated the DKK2 promoter, as revealed by mutation analysis, overexpression and RNA interference (RNAi) experiments. We also confirmed that miR-27a is a negative regulator of the DKK2 gene using miR-27a overexpression and inhibition experiments and mutation analyses. RTCA xCELLigence experiments showed that miR-27a suppressed Chinese hamster ovary (CHO) cell proliferation by down-regulating DKK2 gene expression. Taken together, our findings suggest that C/EBPβ and miR-27a control DKK2 transcription.
authors.affiliation: Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture &Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, PR China.|Hubei Key Laboratory of Animal Embryo Engineering and Molecular Breeding, Hubei Academy of Agriculture Science, Wuhan, 430064, PR China.
authors.forename: Hu
authors.initials: H
authors.lastname: Tao
authors.affiliation: Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture &Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, PR China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture &Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, PR China.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture &Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, PR China.
authors.forename: Panfei
authors.initials: P
authors.lastname: Pang
authors.affiliation: Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture &Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, PR China.
authors.forename: Shanzhi
authors.initials: S
authors.lastname: Cai
authors.affiliation: Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture &Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, PR China.
authors.forename: Jialian
authors.initials: J
authors.lastname: Li
authors.affiliation: Hubei Key Laboratory of Animal Embryo Engineering and Molecular Breeding, Hubei Academy of Agriculture Science, Wuhan, 430064, PR China.
authors.forename: Shuqi
authors.initials: S
authors.lastname: Mei
authors.affiliation: Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture &Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, PR China.|The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, PR China.
authors.forename: Fenge
authors.initials: F
authors.lastname: Li
mesh_terms: D024506:5' Flanking Region; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000378:metabolism*; D016466:CHO Cells; D002522:Chlorocebus aethiops; D003412:Cricetulus; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D013552:Swine; D015533:Transcriptional Activation; D060449:Wnt Signaling Pathway
chemical_list: D022782:CCAAT-Enhancer-Binding Protein-beta; D036341:Intercellular Signaling Peptides and Proteins; D035683:MicroRNAs