Negative Glucocorticoid Response-Like Element from the First Intron of the Chicken Growth Hormone Gene Represses Gene Expression in the Rat Pituitary Tumor Cell Line.
The effects of introns, especially the first intron, on the regulation of gene expression remains unclear. Therefore, the objective of the present study was to investigate the transcriptional regulatory function of intron 1 on thechicken growth hormone(cGH) gene in the rat pituitary tumor cell line (GH4-C1). Transient transfection using first-intron-insertedcGHcomplete coding sequences (CDSs) and non-intron-insertedcGHCDS plasmids, quantitative RT-PCR (qRT-PCR) and western blot assays were used to detect the expression ofcGH. The reporter gene assay was also used to investigate the effect of a series of fragments in the first intron ofcGHon gene expression in GH4-C1. All of the results revealed that a 200-bp fragment located in the +485/+684 region of intron 1 was essential for repressing the expression ofcGH. Further informatics analysis showed that there was a cluster of 13 transcriptional factor binding sites (TFBSs) in the +485/+684 region of thecGHintron 1. Disruption of a glucocorticoid response-like element (the 19-nucleotide sequence 5'-AGGCTTGACAGTGACCTCC-3') containing a T-box motif (TGACCT) located within this DNA fragment increased the expression of the reporter gene in GH4-C1. In addition, an electrophoretic mobility shift assay (EMSA) revealed a glucocorticoid receptor (GR) protein of rat binding to the glucocorticoid response-like element. Together, these results indicate that there is a negative glucocorticoid response-like element (nGRE) located in the +591/+609 region within the first intron ofcGH, which is essential for the down-regulation ofcGHexpression.
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. magiel1018@stu.scau.edu.cn.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. magiel1018@stu.scau.edu.cn.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. magiel1018@stu.scau.edu.cn.
Jing-E
JE
Ma
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. langqianqian@stu.scau.edu.cn.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. langqianqian@stu.scau.edu.cn.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. langqianqian@stu.scau.edu.cn.
Qian-Qian
QQ
Lang
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. qff_1804@163.com.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. qff_1804@163.com.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. qff_1804@163.com.
Feng-Fang
FF
Qiu
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. zhangli761101@163.com.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. zhangli761101@163.com.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. zhangli761101@163.com.
Li
L
Zhang
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. 13751897799@163.com.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. 13751897799@163.com.
Xiang-Guang
XG
Li
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. luowen729@163.com.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. luowen729@163.com.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. luowen729@163.com.
Wen
W
Luo
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. wangjuan901226@126.com.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. wangjuan901226@126.com.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. wangjuan901226@126.com.
Juan
J
Wang
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. wangxing635@163.com.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. wangxing635@163.com.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. wangxing635@163.com.
Xing
X
Wang
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. 15626195391@163.com.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. 15626195391@163.com.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. 15626195391@163.com.
Xi-Ran
XR
Lin
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. wsliu@scau.edu.cn.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. wsliu@scau.edu.cn.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. wsliu@scau.edu.cn.
Wen-Sheng
WS
Liu
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. nqinghua@scau.edu.cn.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. nqinghua@scau.edu.cn.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. nqinghua@scau.edu.cn.
Qing-Hua
QH
Nie
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou 510642, China. xqzhang@scau.edu.cn.|Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, South China Agricultural University, Guangzhou 510642, China. xqzhang@scau.edu.cn.|Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou 510642, China. xqzhang@scau.edu.cn.
Xi-Quan
XQ
Zhang
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D045744:Cell Line, Tumor; D002645:Chickens; D005091:Exons; D005786:Gene Expression Regulation; D005938:Glucocorticoids / Q000494:pharmacology; D013006:Growth Hormone / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007438:Introns; D016366:Open Reading Frames; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000235:genetics* / Q000378:metabolism; D020218:Response Elements; D052683:Somatotrophs / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection; D019076:Transgenes
D005938:Glucocorticoids; D011965:Receptors, Glucocorticoid; D014157:Transcription Factors; D013006:Growth Hormone
chicken growth hormone gene; gene expression; intron 1