文献类型：期刊文献

中文题名：Identification and characterization of the minimal androgen-regulated kidney-specific kidney androgen-regulated protein gene promoter

英文题名：Identification and characterization of the minimal androgen-regulated kidney-specific kidney androgen-regulated protein gene promoter

作者：Liqiang Fan[1,2];Dianne O Hardy[2];James F Catteral[2];Jian Zhao[1];Suxia Li[1]

机构：[1]The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China;[2]Centre for Biomedical Research, The Population Council, New York 10021, USA

年份：2008

卷号：40

期号：12

起止页码：979

中文期刊名：Acta Biochimica et Biophysica Sinica

外文期刊名：生物化学与生物物理学报（英文版）

收录：CSTPCD;;Scopus;北大核心:【北大核心2004】；CSCD:【CSCD2011_2012】；PubMed;

语种：英文

中文关键词：androgen receptor; kidney; proximal tubule cell; promoter

外文关键词：androgen receptor; kidney; proximal tubule cell; promoter

摘要：The kidney androgen-regulated protein （Kap） gene is tissue specific and regulated by androgen in mouse kidney proximal tubule cells （PTCs）. In the present study, we aimed to identify the minimal PTC-specific androgen-regulated Kap promoter and analyze its androgen response elements （AREs）. A deletion series of the Kap1542 promoter/luciferase constructs were assayed in opossum kidney （OK） PTCs in the presence or absence of 15 nM dihydrotestosterone （DHT）. Kap1542 and Kap637 had low activity and no androgen induction; Kap224 had a basal activity that was 4- to 5-fold higher than that of Kap 1542, but was only slightly induced by DHT. Kap147 had a basal activity that was 2- to 3-fold higher than that of Kap1542 and was induced by DHT 4- to 6-fold. Kap77 abolished basal promoter activity but was still induced by DHT. Results showed that, in vitro, Kap147 was a minimal androgen-regulated promoter. Transient transfection in different cells demonstrated that Kap147 specifically initiated reporter gene expression in PTCs. Sequence analysis revealed two potential AREs located at positions -124 and -39 of Kap 147. Mutational assays showed that only the ARE at -124 was involved in androgen response in OK cells. Electrophoretic mobility shift assay also verified -124 ARE bound specifically to androgen receptor. In conclusion, we defined the minimal Kap147 promoter that may be a good model for the study of kidney PTC-specific expression and molecular mechanisms that lead to an androgen-specific responsiveness in vivo.
The kidney androgen-regulated protein （Kap） gene is tissue specific and regulated by androgen in mouse kidney proximal tubule cells （PTCs）. In the present study, we aimed to identify the minimal PTC-specific androgen-regulated Kap promoter and analyze its androgen response elements （AREs）. A deletion series of the Kap1542 promoter/luciferase constructs were assayed in opossum kidney （OK） PTCs in the presence or absence of 15 nM dihydrotestosterone （DHT）. Kap1542 and Kap637 had low activity and no androgen induction; Kap224 had a basal activity that was 4- to 5-fold higher than that of Kap 1542, but was only slightly induced by DHT. Kap147 had a basal activity that was 2- to 3-fold higher than that of Kap1542 and was induced by DHT 4- to 6-fold. Kap77 abolished basal promoter activity but was still induced by DHT. Results showed that, in vitro, Kap147 was a minimal androgen-regulated promoter. Transient transfection in different cells demonstrated that Kap147 specifically initiated reporter gene expression in PTCs. Sequence analysis revealed two potential AREs located at positions -124 and -39 of Kap 147. Mutational assays showed that only the ARE at -124 was involved in androgen response in OK cells. Electrophoretic mobility shift assay also verified -124 ARE bound specifically to androgen receptor. In conclusion, we defined the minimal Kap147 promoter that may be a good model for the study of kidney PTC-specific expression and molecular mechanisms that lead to an androgen-specific responsiveness in vivo.