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MEF 2 and NR 2F2 cooperate to regulate Akr1c14 gene expression in mouse MA ‐10 Leydig cells
Author(s) -
DiLuoffo M.,
Brousseau C.,
Tremblay J. J.
Publication year - 2016
Publication title -
andrology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.947
H-Index - 43
eISSN - 2047-2927
pISSN - 2047-2919
DOI - 10.1111/andr.12150
Subject(s) - mef2 , transcription factor , promoter , chemistry , cyp17a1 , biology , microbiology and biotechnology , gene expression , gene , biochemistry , enhancer
Summary Leydig cells are essential for male reproductive development and health throughout life. Production of androgens [testosterone, dihydrotestosterone ( DHT )] as well as intermediate steroids [progesterone, dihydroprogesterone ( DHP )] is tightly regulated. In the mouse, the 3α‐hydroxysteroid dehydrogenase enzyme (3α‐ HSD , AKR 1C14) catalyses the interconversion of DHP and DHT into less potent steroids. Despite its importance, nothing is currently known regarding the regulation of Akr1c14 expression in Leydig cells. Recently, the transcription factors MEF 2 and NR 2F2 were identified in the mouse testis including in Leydig cells where they were found to regulate expression of genes involved in steroidogenesis. Analyses of transcriptomic data from MEF 2‐ or NR 2F2‐deficient MA ‐10 Leydig cells revealed a significant decrease in Akr1c14 mRNA levels. Using qPCR , we confirmed that Akr1c14 mRNA levels were decreased in MEF 2‐ and in NR 2F2‐deficient conditions. Conversely, overexpression of MEF 2A or/and NR 2F2 in MA ‐10 Leydig cells led to an increase in endogenous Akr1c14 mRNA levels. Recruitment of MEF 2 and NR 2F2 to the Akr1c14 promoter was confirmed by Ch IP while DNA precipitation assays revealed direct binding of MEF 2 but not NR 2F2 to this region. In functional promoter studies, NR 2F2 was found to activate the Akr1c14 promoter while MEF 2A on its own had no effect. Combination of both NR 2F2 and MEF 2A led to a cooperative activation of the Akr1c14 promoter and this required intact MEF 2 and NR 2F2 elements. Finally, co‐immunoprecipitation experiments showed that MEF 2 and NR 2F2 are present in the same protein complex. In conclusion, our results identify a novel cooperation between MEF 2 factors and NR 2F2 in the expression of the Akr1c14 gene involved in the regulation of DHP / DHT levels.