Identification and characterization of the minimal androgen‐regulated kidney‐specific kidney androgen‐regulated protein gene promoter

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 deletio...

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Published in:Acta biochimica et biophysica Sinica 2008-12, Vol.40 (12), p.979-988
Main Authors: Fan, Liqiang, Hardy, Dianne O, Catteral, James F, Zhao, Jian, Li, Suxia
Format: Article
Language:English
Subjects:
androgen receptor
Androgens - physiology
Animals
Base Sequence
Cell Line
DNA Primers
Electrophoretic Mobility Shift Assay
kidney
Mice
Mutagenesis, Site-Directed
promoter
Promoter Regions, Genetic
Proteins - genetics
proximal tubule cell
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Summary: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 Kap1542, 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 Kap147. 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.
ISSN:1672-9145
1745-7270
DOI:10.1111/j.1745-7270.2008.00482.x