This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ris-Stalpers, C. Articles by Brinkmann, A. O. Search for Related Content PubMed PubMed Citation Articles by Ris-Stalpers, C. Articles by Brinkmann, A. O.

Substitution of Aspartic Acid-686 by Histidine or Asparagine in the Human Androgen Receptor Leads to a Functionally Inactive Protein with Altered Hormone-Binding Characteristics

Department of Endocrinology and Reproduction, Erasmus University Rotterdam, The Netherlands
Department of Pathology, Erasmus University Rotterdam, The Netherlands
Lady Davis Institute, Sir M. B. Davis-Jewish General Hospital, Departments of Biology, Medicine, and Pediatrics, McGill University Montreal, Quebec, Canada
Department of Internal Medicine, University of Bonn Bonn, Germany
Ben May Institute, Departments of Biochemistry, Molecular Biology, and Pediatrics, University of Chicago Chicago, Illinois 60637

Address requests for reprints to: Dr. C. Ris-Stalpers, Department of Endocrinology and Reproduction, Erasmus University Rotterdam, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands.

Abstract

We have identified two different single nucleotide alterations in codon 686 (GAC; aspartic acid) in exon 4 of the human androgen receptor gene in three unrelated families with the complete form of androgen insensitivity. One mutation (G C) results in an aspartic acid histidine substitution (with 15–20% of wild-type androgen-binding capacity), whereas the other mutation (G A) leads to an aspartic acid asparagine substitution (with normal androgenbinding capacity, but a rapidly dissociating ligandreceptor complex). The mutations eliminate a Hinfl restriction site. Screening for the loss of the Hinfl site in both families with the Asp Asn mutation resulted in the recognition of heterozygous carriers in successive generations of each. Both mutant androgen receptors were generated in vitro and transiently expressed in COS and HeLa cells. The receptor proteins produced had the same altered binding characteristics as those measured in fibroblasts from the affected subjects. R1881-activated tran-scription of a GRE-tk-CAT reporter gene construct was strongly diminished by both mutant receptors and was only partially restored using a 100-fold higher concentration of ligand compared with wildtype receptor. Thus, aspartic acid-686 appears essential for normal androgen receptor function. Substitution of this amino acid residue, by either histidine or asparagine, results in androgen insensitivity and lack of androgen-dependent male sexual differentiation.

FOOTNOTES

This work was supported by The Netherlands Organization for Scientific Research (to C.R., G.J., J.T., and A.O.B.), the Medical Research Council of Canada Group Grant in Medical Genetics (to M.T., M.K., and L.P.), NIH Grants DK-37694 (to S.L.) and HD-06308 (to R.L.R.), and the Deutsche Forschungsgemeinschaft (Grant Schw 168/5–9).

Received for publication February 21, 1991. Revision received July 29, 1991. Accepted for publication July 29, 1991.

This article has been cited by other articles:

				S. E. Hannema, I. S. Scott, J. Hodapp, H. Martin, N. Coleman, J. W. Schwabe, and I. A. Hughes Residual Activity of Mutant Androgen Receptors Explains Wolffian Duct Development in the Complete Androgen Insensitivity Syndrome J. Clin. Endocrinol. Metab., November 1, 2004; 89(11): 5815 - 5822. [Abstract] [Full Text] [PDF]

				N. Poujol, S. Lumbroso, B. Terouanne, J.-M. A. Lobaccaro, W. Bourguet, and C. Sultan Pathophysiology of Androgen Insensitivity Syndromes: Molecular and Structural Approaches of Natural and Engineered Androgen Receptor Mutations at Amino Acid 743 J. Clin. Endocrinol. Metab., December 1, 2002; 87(12): 5793 - 5800. [Abstract] [Full Text] [PDF]

				S. Rosa, A. Biason-Lauber, N. P. Mongan, F. Navratil, and E. J. Schoenle Complete Androgen Insensitivity Syndrome Caused by a Novel Mutation in the Ligand-Binding Domain of the Androgen Receptor: Functional Characterization J. Clin. Endocrinol. Metab., September 1, 2002; 87(9): 4378 - 4382. [Abstract] [Full Text] [PDF]

				M. J. McPhaul Molecular Defects of the Androgen Receptor Recent Prog. Horm. Res., January 1, 2002; 57(1): 181 - 194. [Abstract] [Full Text] [PDF]

				M. W. Elmlinger, I. Mayer, D. Schnabel, B. S. Schuett, D. Diesing, G. Romalo, H. A. Wollmann, W. Weidemann, K.-D. Spindler, M. B. Ranke, et al. Decreased Expression of IGF-II and Its Binding Protein, IGF-Binding Protein-2, in Genital Skin Fibroblasts of Patients with Complete Androgen Insensitivity Syndrome Compared with Normally Virilized Males J. Clin. Endocrinol. Metab., October 1, 2001; 86(10): 4741 - 4746. [Abstract] [Full Text] [PDF]

				P. A. Koivisto, J. Schleutker, H. Helin, C. E.-v. Eekelen, O.-P. Kallioniemi, and J. Trapman Androgen Receptor Gene Alterations and Chromosomal Gains and Losses in Prostate Carcinomas Appearing During Finasteride Treatment for Benign Prostatic Hyperplasia Clin. Cancer Res., November 1, 1999; 5(11): 3578 - 3582. [Abstract] [Full Text] [PDF]

				M. J. McPhaul and J. E. Griffin Male Pseudohermaphroditism Caused by Mutations of the Human Androgen Receptor J. Clin. Endocrinol. Metab., October 1, 1999; 84(10): 3435 - 3441. [Full Text] [PDF]

				D. L. Shkolny, L. K. Beitel, J. Ginsberg, G. Pekeles, L. Arbour, L. Pinsky, and M. A. Trifiro Discordant Measures of Androgen-Binding Kinetics in Two Mutant Androgen Receptors Causing Mild or Partial Androgen Insensitivity, Respectively J. Clin. Endocrinol. Metab., February 1, 1999; 84(2): 805 - 810. [Abstract] [Full Text]

				H. T. Bruggenwirth, A. L. M. Boehmer, J. M. Lobaccaro, L. Chiche, C. Sultan, J. Trapman, and A. O. Brinkmann Substitution of Ala564 in the First Zinc Cluster of the Deoxyribonucleic Acid (DNA)-Binding Domain of the Androgen Receptor by Asp, Asn, or Leu Exerts Differential Effects on DNA Binding Endocrinology, January 1, 1998; 139(1): 103 - 110. [Abstract] [Full Text] [PDF]