Author:

Keywords:

Aldehyde Reductase, Androgens, Animals, Base Sequence, Cell Line, DNA-Binding Proteins, Humans, Male, Mice, Molecular Sequence Data, Mutagenesis, Site-Directed, Mutation, NFI Transcription Factors, Promoter Regions (Genetics), Proteins, Research Support, Non-U.S. Gov't, Sp1 Transcription Factor, Trans-Activation (Genetics), Transcription Factors, Transcription, Genetic, Transfection, Science & Technology, Life Sciences & Biomedicine, Cell Biology, Endocrinology & Metabolism, androgen response element, transcriptional regulation, transcription factor, NF1, Sp1, mouse vas deferens protein, mutagenesis, transient transfection, MAMMARY-TUMOR VIRUS, NUCLEAR FACTOR-I, KETO REDUCTASE SUPERFAMILY, MMTV PROMOTER, RESPONSE ELEMENT, GENE-EXPRESSION, DNA-BINDING, HORMONE RECEPTORS, MAJOR PROTEIN, C3(1) GENE, Promoter Regions, Genetic, Transcriptional Activation, 06 Biological Sciences, 07 Agricultural and Veterinary Sciences, 11 Medical and Health Sciences, 3101 Biochemistry and cell biology, 3105 Genetics, 3202 Clinical sciences

Abstract:

Transcription of the mouse vas deferens protein (MVDP) gene is stimulated by androgens and we have previously shown that in a 162 bp fragment, located at position -121 to +41, a TGAAGTtccTGTTCT sequence functions as an androgen-dependent enhancer. To determine which factors are involved in the hormonally regulated MVDP gene transcription, we have used DNase I footprinting and band-shift assays to examine in vitro binding of proteins to the enhancer and promoter sequences and have determined the functional significance of the recognized DNA sequences in transient transfection assays. Studies using recombinant proteins such as the DNA binding domain of the androgen receptor (AR-DBD) and Sp1, and crude cellular extracts from T47D and vas deferens epithelial cells (VDEC) showed that in addition to AR-DBD, the transcriptional activators NF1 and Sp1 interact with the -121/+41 fragment in a specific manner. Transient transfection assays revealed that site-directed mutations in the NF1 and Sp1 binding elements strongly reduced (NF1) or abolished (Sp1) androgen induced expression. The results demonstrate that the -121/+41 sequence is a composite site for the androgen receptor mediated transactivation of the MVDP gene.