Regulatory capacity of an androgen-specific enhancer of the mouse Slp gene in transgenic mice.
Different steroid hormone receptors can activate transcription from the same hormone response element (HRE) in vitro, but in vivo the effects of each hormone on gene activity are distinct. To determine sequences mediating androgen-specific response in a physiological setting, we placed the androgen-responsive mouse sex-limited protein gene (Slp) enhancer before a tkCAT reporter in transgenic mice. The enhancer contains a consensus HRE plus accessory factor binding sites that act in concert to direct transcription in response to androgen. A 160 bp fragment, C'delta2, is responsive to several steroids in transfection; in transgenic mice, this enhancer was active in several tissues of male and female mice, in four of six transgenic lines. In striking contrast, C'delta9, a 120 bp sub-fragment of C'delta2 that responds only to androgen in transfection, showed activity in testes, prostate and kidney, where it was strongly androgen-inducible in females. However, expression was obtained in only one transgenic line. Multimerization of the C'delta9 enhancer conferred expression in prostate, but again in only one line. The greater penetrance of C'delta2 expression was not driven by glucocorticoids, as adrenalectomy had little effect, but may be dependent on the NF-kappaB-like element absent from the C'delta9 fragment. That two transgenic lines showed expression in androgen target sites driven by enhancers that are androgen-specific in vitro suggested that activation of this enhancer, when it could occur, was in response to androgen. The dramatically different behavior of the two related enhancer sequences underscores the importance of chromosomal context to the activity and specificity of regulatory elements.
Nelson
S A
SA
Department of Human Genetics, University of Michigan Medical School, Ann Arbor 48109-0618, USA.
Robins
D M
DM
D000311:Adrenal Glands / Q000502:physiology; D000315:Adrenalectomy; D000728:Androgens / Q000502:physiology*; D000818:Animals; D001798:Blood Proteins / Q000235:genetics*; D003181:Complement C4; D003907:Dexamethasone / Q000494:pharmacology; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005260:Female; D005938:Glucocorticoids / Q000494:pharmacology; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009928:Organ Specificity; D011993:Recombinant Fusion Proteins; D013739:Testosterone / Q000494:pharmacology; D015533:Transcriptional Activation / Q000187:drug effects / Q000235:genetics*
D000728:Androgens; D001798:Blood Proteins; C018386:C4a protein, mouse; D003181:Complement C4; D005938:Glucocorticoids; D011993:Recombinant Fusion Proteins; D013739:Testosterone; D003907:Dexamethasone