A mutation inactivating the distal SF1 binding site on the human anti-Müllerian hormone promoter causes persistent Müllerian duct syndrome

Abstract

The persistent Müllerian duct syndrome (PMDS) is a 46,XY disorder of sexual development characterized by the persistence of Müllerian duct derivatives, uterus and tubes, in otherwise normally masculinized males. The condition, transmitted as a recessive autosomal trait, is usually due to mutations in either the anti-Müllerian hormone (AMH) gene or its main receptor. Many variants of these genes have been described, all targeting the coding sequences. We report the first case of PMDS due to a regulatory mutation. The AMH promoter contains two binding sites for steroidogenic factor 1 (SF1), one at −102 and the other at −228. Our patient carries a single base deletion at −225, significantly decreasing its capacity for binding SF1, as measured by the electrophoresis mobility shift assay. Furthermore, by linking the AMH promoter to the luciferase gene, we show that the transactivation capacity of the promoter is significantly decreased by the mutation, in contrast to the disruption of the −102 binding site. To explain the difference in impact we hypothesize that SF1 could partially overcome the lack of binding to the −102 binding site by interacting with a GATA4 molecule linked to a nearby response element. We show that disruption of both the −102 SF1 and the −84 GATA response elements significantly decreases the transactivation capacity of the promoter. In conclusion, we suggest that the distance between mutated SF1 sites and potentially rescuing GATA binding motifs might play a role in the development of PMDS.

Molecular mechanisms underlying AMH elevation in hyperoestrogenic states in males

“…The plasmid of the h AMH promoter with the mutated ERE half-site was generated using the QuikChange II XL Targeted Mutagenesis Kit (Stratagene), as described 15 . Oligonucleotide primers for mutagenesis (Supplementary Table S2 ) were synthesised by Eurogentec (Liège, Belgium).…”

“…In the fetal Sertoli cell, AMH gene expression is triggered by the nuclear transcription factor SOX9 11 , 12 and subsequently upregulated by transcription factors SF1 11 , 13 – 15 , GATA4 14 , 16 , 17 and WT1 18 . Thus, high levels of AMH are secreted during the period of sex differentiation independently of pituitary gonadotrophin action on Sertoli cells.…”

GATA4 regulates epithelial morphogenesis in the developing mouse stomach to promote establishment of a glandular columnar epithelium

“…Among these were several transcription factors that act with GATA factors to direct gene expression programs in other developmental systems including the heart, liver, and neural systems (Maitra et al 2009;Nadeau et al 2010;Ang et al 2016;Watanabe et al 2000;Tremblay and Viger 2001;Viger et al 2008;Schteingart et al 2019;Fokko van Loo et al 2007). For example, TBX5 and GATA4 co-bind genes in cardiac cells to activate gene expression (Maitra et al 2009;Nadeau et al 2010;Ang et al 2016).…”

“…For example, TBX5 and GATA4 co-bind genes in cardiac cells to activate gene expression (Maitra et al 2009;Nadeau et al 2010;Ang et al 2016). SF1 and GATA4 function cooperatively to activate gene expression pathways in Sertoli cells (Watanabe et al 2000;Tremblay and Viger 2001;Viger et al 2008;Schteingart et al 2019). Although SP3, like GATA4, can function to activate or repress gene expression (Majello et al 1997;Lania et al1997), GATA4 and SP3 can interact to activate Carp1 expression in the heart (Fokko van Loo et al 2007).…”

“…We identified different sets of consensus transcription factor binding motifs statistically enriched around GATA4 binding sites (+/-100 bp of the peak) in the HSE and FSE gene sets differentially expressed in GATA4 mutants. Several de novo motifs predicted in the GATA4 binding neighborhoods of activated and repressed gene sets identified transcription factors previously shown to cooperate with GATA4 to direct gene regulatory networks in other organ systems including TBX5, SF1, and SP3 (Maitra et al 2009;Nadeau et al 2010;Ang et al 2016;Watanabe et al 2000;Tremblay and Viger 2001;Viger et al 2008;Schteingart et al 2019;Fokko van Loo et al 2007). Others,…”