title: DNA methylation and NF-Y regulate Piwil1 expression during chicken spermatogenesis.
abstract: The P-element induced wimpy testis (Piwi) protein family, a subfamily of the Argonaute protein family, is involved in gene silencing and shows specific expression in spermatogenic cells. To reveal the transcriptional regulatory mechanisms of Piwil1 in chickens, we cloned sequences of the chicken Piwil1 promoter region and performed luciferase reporter and electrophoretic mobility shift assays to analyze the transcriptional activity and identify important transcriptional regulatory elements. The results showed that the region from -90 to -43 in the 5'-flanking region of Piwil1 contains a transcriptional regulatory CCAAT box that was necessary for the transcriptional activity of the Piwil1 promoter. Moreover, the transcription factor nuclear factor Y (NF-Y) was bound to the Piwil1 promoter CCAAT box specifically in germ cells. In addition, bisulfite sequencing to determine the methylation profile of the Piwil1 promoter CpG island in different spermatogenic and non-germ cell populations was performed. Compared with germ cells, non-germ cells showed increased methylation of the promoter region containing the CCAAT box, loss of NF-Y binding, and silencing of the Piwil1 locus. It is demonstrated that the specific expression of Piwil1 in chicken germ cells is regulated by the transcription factor NF-Y and differential CpG island methylation.
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: passioncgb@163.com.
authors.forename: Guobin
authors.initials: G
authors.lastname: Chang
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China; Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China. Electronic address: chenrong_big@163.com.
authors.forename: Rong
authors.initials: R
authors.lastname: Chen
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: herry2800@163.com.
authors.forename: Lu
authors.initials: L
authors.lastname: Xu
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: 26177435@qq.com.
authors.forename: Teng
authors.initials: T
authors.lastname: Ma
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: 434373554@qq.com.
authors.forename: Hongzhi
authors.initials: H
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: 276532497@qq.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: zyang@yzu.edu.cn.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: 1169587200@qq.com.
authors.forename: Zhiteng
authors.initials: Z
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: 583292863@qq.com.
authors.forename: Fang
authors.initials: F
authors.lastname: Wan
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: 1083866231@qq.com.
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Guo
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: 544460951@qq.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Xu
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: wmzhao@yzu.edu.cn.
authors.forename: Wenming
authors.initials: W
authors.lastname: Zhao
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. Electronic address: ghchen@yzu.edu.cn.
authors.forename: Guohong
authors.initials: G
authors.lastname: Chen
mesh_terms: D000818:Animals; D060565:Argonaute Proteins / Q000502:physiology*; D023081:CCAAT-Binding Factor / Q000502:physiology*; D002645:Chickens; D019175:DNA Methylation / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay / Q000662:veterinary; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D008297:Male; D016297:Mutagenesis, Site-Directed / Q000662:veterinary; D011401:Promoter Regions, Genetic / Q000502:physiology; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D013091:Spermatogenesis / Q000502:physiology*
chemical_list: D060565:Argonaute Proteins; D023081:CCAAT-Binding Factor
keywords: Chicken; DNA methylation; Piwil1; Spermatogenesis