■转录因子SOX9是男性性发育和支持细胞分化的关键调节因子。改变的SOX9表达与哺乳动物性发育障碍(DSD)的发病机理有关。然而,关于在性发育过程中控制其转录控制的表观遗传机制的信息有限。
■本研究采用实时PCR(qPCR),免疫荧光(IIF),和染色质免疫沉淀(ChIP)测定,以研究与人和小鼠Sertoli细胞系中SOX9基因转录控制相关的表观遗传机制。为了确定参与SOX9表观遗传控制的特定表观遗传酶,使用siRNA对P300、GCN5和WDR5进行功能测定。
■SOX9的转录激活与活性组蛋白修饰的选择性沉积有关,如H3K4me3和H3K27ac,在其增强子和启动子区域。重要的是,发现组蛋白乙酰转移酶P300在SOX9增强子上显著富集,与H3K27ac和SOX9转录因子共定位。在eSR-A和e-ALDI增强子中,P300的沉默导致SOX9表达降低和H3K27ac水平降低,证明了P300介导的组蛋白乙酰化在SOX9转录激活中的关键作用。有趣的是,另一种组蛋白赖氨酸乙酰转移酶,如GNC5和甲基转移酶,如Trithorax/COMPASS样,也可能在男性性分化中发挥相关作用。
■在SOX9增强剂上通过P300进行组蛋白乙酰化,是控制男性性发育这一重要调节因子转录控制的关键机制。这些发现为性别分化的表观遗传基础和DSD的潜在发病机理提供了重要见解。
UNASSIGNED: The transcription factor SOX9 is a key regulator of male sexual development and Sertoli cell differentiation. Altered SOX9 expression has been implicated in the pathogenesis of disorders of sexual development (DSD) in mammals. However, limited information exists regarding the epigenetic mechanisms governing its transcriptional control during sexual development.
UNASSIGNED: This study employed real-time PCR (qPCR), immunofluorescence (IIF), and chromatin immunoprecipitation (ChIP) assays to investigate the epigenetic mechanisms associated with SOX9 gene transcriptional control in human and mouse Sertoli cell lines. To identify the specific epigenetic enzymes involved in SOX9 epigenetic control, functional assays using siRNAs for P300, GCN5, and WDR5 were performed.
UNASSIGNED: The transcriptional activation of SOX9 was associated with selective deposition of active histone modifications, such as H3K4me3 and H3K27ac, at its enhancer and promoter regions. Importantly, the histone acetyltransferase P300 was found to be significantly enriched at the SOX9 enhancers, co-localizing with the H3K27ac and the SOX9 transcription factor. Silencing of P300 led to decreased SOX9 expression and reduced H3K27ac levels at the eSR-A and e-ALDI enhancers, demonstrating the crucial role of P300-mediated histone acetylation in SOX9 transcriptional activation. Interestingly, another histone lysine acetyltransferases like GNC5 and methyltransferases as the Trithorax/COMPASS-like may also have a relevant role in male sexual differentiation.
UNASSIGNED: Histone acetylation by P300 at SOX9 enhancers, is a key mechanism governing the transcriptional control of this essential regulator of male sexual development. These findings provide important insights into the epigenetic basis of sexual differentiation and the potential pathogenesis of DSDs.