A Practical Approach to the Detection of Androgen Receptor Gene Mutations and Pedigree Analysis in Families with X-Linked Androgen Insensitivity

Ris-Stalpers, C.; Hoogenboezem, Theo; Sleddens, Hein F.B.M.; Verleun-Mooijman, M.C.T.; Degenhart, H. J.; Drop, S. L. S.; Halley, Dicky; Oosterwijk, JC; Hodgins, Malcolm B.; Trapman, Jan; Brinkmann, Albert O.

doi:10.1203/00006450-199408000-00015

Cited by 42 publications

(17 citation statements)

References 19 publications

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“…4), to CaM-agarose. AR in LNCaP cells has a mutation in the ligand-binding domain, which is known to affect the activity, affinity, and responsiveness of AR to various ligands (41)(42)(43)(44)(45). These observations, taken together, suggest that the ligand-binding domain of AR may play an important role in AR interaction with CaM.…”

Section: Discussionmentioning

confidence: 97%

Physical and functional interaction of androgen receptor with calmodulin in prostate cancer cells

Cifuentes

Mataraza

Yoshida

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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Ca 2؉ and calmodulin (CaM) play a critical role in proliferation and viability of a wide variety of cells, including prostate cancer cells. We examined two prostate cancer cell lines, androgen-sensitive LNCaP and androgen-independent PC-3. Proliferation of LNCaP cells was six to eight times more sensitive to the inhibitory effect of the CaM antagonist N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride (W-7) than were PC-3 cells. Because LNCaP cell proliferation is sensitive to stimulation by androgen, we assessed the physical and functional interaction between androgen receptor (AR) and CaM. We observed tight binding of AR to CaM when LNCaP cell extracts were subjected to CaM-affinity column chromatography. AR binding to CaM was Ca 2؉ -dependent and was inhibited by pretreatment of the cell extracts with W-7. Using immunofluorescence staining and confocal microscopy, we demonstrated colocalization of AR and CaM in the nucleus of LNCaP cells. Furthermore, the functional relevance of AR-CaM interactions in intact cells was revealed by the observation that W-7 was as effective as Casodex, an antiandrogen, in blocking AR-regulated expression of prostate-specific antigen in LNCaP cells. AR seems to interact with CaM directly because purified human AR could bind to CaM-agarose, and CaM could be detected in AR-immunoprecipitate prepared from purified soluble proteins. These studies provide direct evidence for physical and functional interaction between AR and CaM and suggest the potential usefulness of CaM antagonists in blocking AR activity in prostate cancer.

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Section: Discussionmentioning

confidence: 97%

Physical and functional interaction of androgen receptor with calmodulin in prostate cancer cells

Cifuentes

Mataraza

Yoshida

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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“…For example, the binding of flutamide, estradiol, and progesterone to the T877A mutant can activate the mutant receptor (39,40). Conversely, mutation of T877 to residues with larger sidechains such as aspartic acid and lysine would be expected to prevent the binding of ligands with any substituent at position 17 of the D-ring, and such mutations have been shown to eliminate androgen binding (41).…”

Section: Binding Of Dhtmentioning

confidence: 99%

Crystallographic structures of the ligand-binding domains of the androgen receptor and its T877A mutant complexed with the natural agonist dihydrotestosterone

Sack

Kish

Wang

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

436

266

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The structures of the ligand-binding domains (LBD) of the wildtype androgen receptor (AR) and the T877A mutant corresponding to that in LNCaP cells, both bound to dihydrotestosterone, have been refined at 2.0 Å resolution. In contrast to the homodimer seen in the retinoid-X receptor and estrogen receptor LBD structures, the AR LBD is monomeric, possibly because of the extended C terminus of AR, which lies in a groove at the dimerization interface. Binding of the natural ligand dihydrotestosterone by the mutant LBD involves interactions with the same residues as in the wildtype receptor, with the exception of the side chain of threonine 877, which is an alanine residue in the mutant. This structural difference in the binding pocket can explain the ability of the mutant AR found in LNCaP cells (T877A) to accommodate progesterone and other ligands that the wild-type receptor cannot.

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“…The F764L mutation was originally identified in a CAIS patient of Dr. G. Costin (Los Angeles, CA) [11], and has also been reported in other patients by two additional groups [19,20]. Biochemical studies in transfected CHO cells demonstrated normal 3 H-DHT K d and B max (the K d was 0.26 nM for the wild type AR and 0.1 nM for mutant F764L, which is not considered a meaningful difference), and abnormally elevated ligand dissociation rate [11].…”

Section: Mutation F764lmentioning

confidence: 75%

Androgen Receptor Mutations Associated with Androgen Insensitivity Syndrome: A High Content Analysis Approach Leading to Personalized Medicine

Szafran

Sun

Hartig

et al. 2011

Advances in Experimental Medicine and Biology

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Androgen insensitivity syndrome (AIS) is a rare disease associated with inactivating mutations of AR that disrupt male sexual differentiation, and cause a spectrum of phenotypic abnormalities having as a common denominator loss of reproductive viability. No established treatment exists for these conditions, however there are sporadic reports of patients (or recapitulated mutations in cell lines) that respond to administration of supraphysiologic doses (or pulses) of testosterone or synthetic ligands. Here, we utilize a novel high content analysis (HCA) approach to study AR function at the single cell level in genital skin fibroblasts (GSF). We discuss in detail findings in GSF from three historical patients with AIS, which include identification of novel mechanisms of AR malfunction, and the potential ability to utilize HCA for personalized treatment of patients affected by this condition.

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A Practical Approach to the Detection of Androgen Receptor Gene Mutations and Pedigree Analysis in Families with X-Linked Androgen Insensitivity

Cited by 42 publications

References 19 publications

Physical and functional interaction of androgen receptor with calmodulin in prostate cancer cells

Physical and functional interaction of androgen receptor with calmodulin in prostate cancer cells

Crystallographic structures of the ligand-binding domains of the androgen receptor and its T877A mutant complexed with the natural agonist dihydrotestosterone

Androgen Receptor Mutations Associated with Androgen Insensitivity Syndrome: A High Content Analysis Approach Leading to Personalized Medicine

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