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Submitted on October 13, 2004
Accepted on July 8, 2005
Institut für Anatomie und Zellbiologie, Justus-Liebig-Universität Giessen (F.X., J.K.), D-35385 Giessen, Germany; Institut für Molekularbiologie und Tumorforschung (A.M.), Philipps-Universität Marburg, D-35033 Marburg, Germany; Institut für Humangenetik und Anthropologie, Klinikum der Friedrich-Schiller-Universität Jena, 07740 Jena (M.P., A.B.)
* To whom correspondence should be addressed. E-mail: joerg.klug{at}anatomie.med.uni-giessen.de.
Human secretoglobin (SCGB) 2A1 (or lipophilin C, lacryglobin, mammaglobin B) is a small protein of unknown function that forms heterodimers with secretoglobin 1D1 (lipophilin A) in tears and is expressed in the prostate. Here we show that SCGB 2A1 is under androgen control in the androgen responsive prostatic cell line LNCaP and can be induced more than 20-fold by dihydrotestosterone. Only 6 h after androgen treatment a strong DNase I hypersensitive site (DHS) is induced in the proximal promoter within chromatin. Within the boundaries of this DHS a minimal 32 bp peculiar dimeric inverted repeat variant GC box (dim-IR-GA box) was found to confer androgen but not glucocorticoid responsiveness in gene transfer experiments. Mutations of both GA boxes that abolish binding of Sp1 and Sp3 also abrogate the androgen response. In an electrophoretic mobility shift assay (EMSA) the DNA binding domain of the androgen receptor (AR) was not able to bind directly to the dim-IR-GA box. But AR is functionally required for the hormone response because induction can be inhibited with the nonsteroidal antagonist bicalutamide. Chromatin immunoprecipitation experiments demonstrated that AR is recruited to the proximal promoter 10 min after androgen treatment. Therefore we propose that SCGB 2A1 represents a new class of androgen target genes that are purely under indirect AR control mediated by DNA-bound Sp factors.
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