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Molecular Endocrinology, Vol 7, 23-36, Copyright © 1993 by Endocrine Society

ARTICLES

Characterization of two cis-acting DNA elements involved in the androgen regulation of the probasin gene

PS Rennie, N Bruchovsky, KJ Leco, PC Sheppard, SA McQueen, H Cheng, R Snoek, A Hamel, ME Bock and BS MacDonald
Department of Cancer Endocrinology, British Columbia Cancer Agency, Vancouver, Canada.

The location and sequence of androgen responsive elements (AREs) in the 5'-flanking DNA of the androgen-regulated rat probasin (PB) gene were determined. The DNA- and steroid-binding domains of the rat androgen receptor [glutathione-S-transferase (GST)-AR1] and the DNA-binding domain and hinge region alone (GST-AR2) were expressed in Escherichia coli as isopropyl-B-D-thioglactopyranoside-induced fusion proteins with GST and purified using glutathione affinity chromatography. Band shift assays indicated that the AR1 peptide was at least five times more effective than AR2 in binding to PB 5'-flanking DNA (-426 to +28), although both gave qualitatively similar patterns and were displaced by anti-AR antibodies. DNase I footprinting experiments revealed two putative AREs: one between positions -236 and -223 (ARE-1) and the other between -140 and -117 (ARE-2). Hormonal regulation of PB was determined by cotransfecting reporter constructions containing the PB 5'-flanking region (-426 to +28) linked to the bacterial chloramphenicol acetyl transferase (CAT) gene with androgen, glucocorticoid, or progesterone receptor expression vectors into human prostatic carcinoma cells (PC-3). PB-CAT gene expression was more effectively induced by androgens than by glucocorticoids or progestins. Both 5'- and 3'-deletion mapping of the PB 5'-flanking DNA revealed that ARE-1 and ARE-2 were required for androgen regulation. A single base mutation in either ARE resulted in a more than 95% loss of androgen induction of CAT. In comparable transfection experiments, the PB hormone-responsive elements showed a greater induction by androgens than did mouse mammary tumor virus or tyrosine aminotransferase elements. Thus, the preferential androgen regulation of the PB gene involves the participation of two different cis-acting DNA elements that bind AR.


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C. S. Song, M. H. Jung, S. C. Kim, T. Hassan, A. K. Roy, and B. Chatterjee
Tissue-specific and Androgen-repressible Regulation of the Rat Dehydroepiandrosterone Sulfotransferase Gene Promoter
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S. Thornton, D. W. Thomas, P. M. Gallagher, and R. E. Ganschow
Androgen Responsiveness of Mouse Kidney {beta}-Glucuronidase Requires 5'-Flanking and Intragenic Gus-s Sequences
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C. B. J. M. Cleutjens, K. Steketee, C. C. E. M. van Eekelen, J. A. G. M. van der Korput, A. O. Brinkmann, and J. Trapman
Both Androgen Receptor and Glucocorticoid Receptor Are Able to Induce Prostate-Specific Antigen Expression, but Differ in Their Growth-Stimulating Properties of LNCaP Cells
Endocrinology, December 1, 1997; 138(12): 5293 - 5300.
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Z. Wang, R. Tufts, R. Haleem, and X. Cai
Genes regulated by androgen in the rat ventral prostate
PNAS, November 25, 1997; 94(24): 12999 - 13004.
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 EndocrinologyHome page
J.-D. Shan, K. Porvari, M. Ruokonen, A. Karhu, V. Launonen, P. Hedberg, J. Oikarinen, and P. Vihko
Steroid-Involved Transcriptional Regulation of Human Genes Encoding Prostatic Acid Phosphatase, Prostate-Specific Antigen, and Prostate-Specific Glandular Kallikrein
Endocrinology, September 1, 1997; 138(9): 3764 - 3770.
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N. Sato, M. D. Sadar, N. Bruchovsky, F. Saatcioglu, P. S. Rennie, S. Sato, P. H. Lange, and M. E. Gleave
Androgenic Induction of Prostate-specific Antigen Gene Is Repressed by Protein-Protein Interaction between the Androgen Receptor and AP-1/c-Jun in the Human Prostate Cancer Cell Line LNCaP
J. Biol. Chem., July 11, 1997; 272(28): 17485 - 17494.
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M. C. W. Avellar, C. W. Gregory, S. G. A. Power, and F. S. French
Androgen-dependent Protein Interactions within an Intron 1 Regulatory Region of the 20-kDa Protein Gene
J. Biol. Chem., July 11, 1997; 272(28): 17623 - 17631.
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A. Devos, F. Claessens, P. Alen, J. Winderickx, W. Heyns, W. Rombauts, and B. Peeters
Identification of a Functional Androgen-Response Element in the Exon 1-Coding Sequence of the Cystatin-Related Protein Gene crp2
Mol. Endocrinol., July 1, 1997; 11(8): 1033 - 1043.
[Abstract] [Full Text]

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 EndocrinologyHome page
S. S. Lim-Tio, M.-C. Keightley, and P. J. Fuller
Determinants of Specificity of Transactivation by the Mineralocorticoid or Glucocorticoid Receptor
Endocrinology, June 1, 1997; 138(6): 2537 - 2543.
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Z. Zhou, J. L. Corden, and T. R. Brown
Identification and Characterization of a Novel Androgen Response Element Composed of a Direct Repeat
J. Biol. Chem., March 28, 1997; 272(13): 8227 - 8235.
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K. B. J. M. Cleutjens, H. A. G. M. van der Korput, C. C. E. M. van Eekelen, H. C. J. van Rooij, P. W. Faber, and J. Trapman
An Androgen Response Element in a Far Upstream Enhancer Region Is Essential for High, Androgen-Regulated Activity of the Prostate-Specific Antigen Promoter
Mol. Endocrinol., February 1, 1997; 11(2): 148 - 161.
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M. Burgos-Trinidad, G. L. Youngblood, M. R. Maroto, A. Scheller, D. M. Robins, and A. H. Payne
Repression of cAMP-Induced Expression of The Mouse P450 17{{alpha}}-Hydroxylase/C17-20 Lyase Gene (Cyp17) by Androgens
Mol. Endocrinol., January 1, 1997; 11(1): 87 - 96.
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S. E. Fenton, N. S. Groce, and D. C. Lee
Characterization of the Mouse Epidermal Growth Factor Promoter and 5'-Flanking Region. ROLE FOR AN ATYPICAL TATA SEQUENCE
J. Biol. Chem., November 29, 1996; 271(48): 30870 - 30878.
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E. T. Keller, C. Chang, and W. B. Ershler
Inhibition of NFkappa B Activity through Maintenance of Ikappa Balpha Levels Contributes to Dihydrotestosterone-mediated Repression of the Interleukin-6 Promoter
J. Biol. Chem., October 18, 1996; 271(42): 26267 - 26275.
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L. V. Nazareth and N. L. Weigel
Activation of the Human Androgen Receptor through a Protein Kinase A Signaling Pathway
J. Biol. Chem., August 16, 1996; 271(33): 19900 - 19907.
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F. Claessens, P. Alen, A. Devos, B. Peeters, G. Verhoeven, and W. Rombauts
The Androgen-specific Probasin Response Element 2Interacts Differentially with Androgen and Glucocorticoid Receptors
J. Biol. Chem., August 9, 1996; 271(32): 19013 - 19016.
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K. B. J. M. Cleutjens, C. C. E. M. van Eekelen, H. A. G. M. van der Korput, A. O. Brinkmann, and J. Trapman
Two Androgen Response Regions Cooperate in Steroid Hormone Regulated Activity of the Prostate-specific Antigen Promoter
J. Biol. Chem., March 15, 1996; 271(11): 6379 - 6388.
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E. Langley, Z.-x. Zhou, and E. M. Wilson
Evidence for an Anti-parallel Orientation of the Ligand-activated Human Androgen Receptor Dimer
J. Biol. Chem., December 15, 1995; 270(50): 29983 - 29990.
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H. H. Juang, L. C. Costello, and R. B. Franklin
Androgen Modulation of Multiple Transcription Start Sites of the Mitochondrial Aspartate Aminotransferase Gene in Rat Prostate
J. Biol. Chem., May 26, 1995; 270(21): 12629 - 12634.
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B. He, J. A. Kemppainen, and E. M. Wilson
FXXLF and WXXLF Sequences Mediate the NH2-terminal Interaction with the Ligand Binding Domain of the Androgen Receptor
J. Biol. Chem., July 21, 2000; 275(30): 22986 - 22994.
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G. Han, B. A. Foster, S. Mistry, G. Buchanan, J. M. Harris, W. D. Tilley, and N. M. Greenberg
Hormone Status Selects for Spontaneous Somatic Androgen Receptor Variants That Demonstrate Specific Ligand and Cofactor Dependent Activities in Autochthonous Prostate Cancer
J. Biol. Chem., March 30, 2001; 276(14): 11204 - 11213.
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M. I. Gonzalez and D. M. Robins
Oct-1 Preferentially Interacts with Androgen Receptor in a DNA-dependent Manner That Facilitates Recruitment of SRC-1
J. Biol. Chem., February 23, 2001; 276(9): 6420 - 6428.
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K. J. Reid, S. C. Hendy, J. Saito, P. Sorensen, and C. C. Nelson
Two Classes of Androgen Receptor Elements Mediate Cooperativity through Allosteric Interactions
J. Biol. Chem., January 19, 2001; 276(4): 2943 - 2952.
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 Proc. Natl. Acad. Sci. USAHome page
M. Stanbrough, I. Leav, P. W. L. Kwan, G. J. Bubley, and S. P. Balk
Prostatic intraepithelial neoplasia in mice expressing an androgen receptor transgene in prostate epithelium
PNAS, September 11, 2001; 98(19): 10823 - 10828.
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