Department of Pediatrics, University of North Carolina, Chapel Hill, North Carolina 27599-7500, USA.
J Biol Chem. 2012 Mar 30;287(14):10905-15. doi: 10.1074/jbc.M111.336081. Epub 2012 Feb 13.
Naturally occurring germ line mutations in the X-linked human androgen receptor (AR) gene cause incomplete masculinization of the external genitalia by disrupting AR function in males with androgen insensitivity syndrome. Almost all AR missense mutations that cause androgen insensitivity syndrome are located in the highly structured DNA and ligand binding domains. In this report we investigate the functional defect associated with an AR exon 1 missense mutation, R405S, that caused partial androgen insensitivity. The 46,XX heterozygous maternal carrier had a wild-type Arg-405 CGC allele but transmitted an AGC mutant allele coding for Ser-405. At birth, the 46,XY proband had a bifid scrotum, hypospadias, and micropenis consistent with clinical stage 3 partial androgen insensitivity. Androgen-dependent transcriptional activity of AR-R405S expressed in CV1 cells was less than wild-type AR and refractory in androgen-dependent AR NH(2)- and carboxyl interaction transcription assays that depend on the coregulator effects of melanoma antigen-A11. This mutation created a Ser-405 phosphorylation site evident by the gel migration of an AR-R405S NH(2)-terminal fragment as a double band that converted to the wild-type single band after treatment with λ-phosphatase. Detrimental effects of the R405S mutation were related to the proximity of the AR WXXLF motif (433)WHTLF(437) required for melanoma antigen-A11 and p300 to stimulate transcriptional activity associated with the AR NH(2)- and carboxyl-terminal interaction. We conclude that the coregulator effects of melanoma antigen-A11 on the AR NH(2)- and carboxyl-terminal interaction amplify the androgen-dependent transcriptional response to p300 required for normal human male sex development in utero.
X 连锁人类雄激素受体 (AR) 基因中的种系突变导致雄激素不敏感综合征男性的 AR 功能障碍,从而导致外生殖器不完全男性化。几乎所有导致雄激素不敏感综合征的 AR 错义突变都位于高度结构化的 DNA 和配体结合结构域中。在本报告中,我们研究了与 AR 外显子 1 错义突变 R405S 相关的功能缺陷,该突变导致部分雄激素不敏感。46,XX 杂合母体携带者具有野生型 Arg-405 CGC 等位基因,但传递了编码 Ser-405 的 AGC 突变等位基因。出生时,46,XY 先证者具有双叉阴囊、尿道下裂和小阴茎,符合部分雄激素不敏感的临床 3 期。CV1 细胞中表达的 AR-R405S 的雄激素依赖性转录活性低于野生型 AR,并且在雄激素依赖性 AR NH(2)-和羧基相互作用转录测定中无反应,该测定依赖于黑色素瘤抗原-A11 的共调节剂效应。该突变创建了一个 Ser-405 磷酸化位点,通过 AR-R405S NH(2)-末端片段的凝胶迁移可见,该片段为双带,在用 λ-磷酸酶处理后转换为野生型单带。R405S 突变的有害影响与 AR WXXLF 基序 (433)WHTLF(437) 有关,该基序需要黑色素瘤抗原-A11 和 p300 来刺激与 AR NH(2)-和羧基末端相互作用相关的转录活性。我们得出结论,黑色素瘤抗原-A11 对 AR NH(2)-和羧基末端相互作用的共调节剂效应放大了 p300 对正常人类男性性发育所必需的雄激素依赖性转录反应。