FOXL2 transcriptionally represses Sf1 expression by antagonizing WT1 during ovarian development in mice
FOXL2 transcriptionally represses Sf1 expression by antagonizing WT1 during ovarian development in mice
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DOI：
10.1096/fj.13-246108
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发表时间：
2014-05-01
期刊：
影响因子：
4.8
通讯作者：
Koopman, Peter
中科院分区：
文献类型：
--
作者：
Takasawa, Kei;Kashimada, Kenichi;Koopman, Peter
类固醇生成因子1（Steroidogenicfactor 1，SF 1; Ad 4 BP/NR 5A 1）在性腺发育中起重要作用。最初，Sf 1基因在两种性别的小鼠胎儿性腺中表达，但后来在睾丸中上调，在卵巢中下调。虽然Sf 1的表达被Wilms tumor 1（WT 1）和LIM homeobox 9（LHX 9）激活并维持，但性别特异性调控的机制仍不清楚。我们假设Sf 1在卵巢发育过程中受到转录因子叉头盒L2（FOXL 2）的抑制。在体外系统（TM 3细胞）中，通过定量RT-PCR测定，FOXL 2拮抗WT 1剪接变体WT 1-KTS对Sf 1的上调。使用报告分析，我们本地化的Sf 1近端启动子区域参与这种拮抗作用的674 bp的间隔。一个保守的FOXL 2结合位点被确定在这段时间内，在体外染色质免疫沉淀。在报告基因测定中，将突变引入该位点消除了FOXL 2的负调控。最后，在Foxl 2基因敲除小鼠中，Sf 1的表达相对于野生型XX胎儿性腺增加了2倍。我们的结果支持FOXL 2在小鼠卵巢发育早期通过拮抗WT 1-KTS负调控Sf 1表达的假设。Kashimada，K.，佩洛西，E.，Takagi，M.，Morio，T.，Asahara，H.，Schlessinger，D.，Mizutani，S.，Koopman，P. FOXL 2在小鼠卵巢发育过程中通过拮抗WT 1转录抑制Sf 1表达。
Steroidogenic factor 1 (SF1; Ad4BP/NR5A1) plays key roles in gonadal development. Initially, the Sf1 gene is expressed in mouse fetal gonads of both sexes, but later is up-regulated in testes and down-regulated in ovaries. While Sf1 expression is activated and maintained by Wilms tumor 1 (WT1) and LIM homeobox 9 (LHX9), the mechanism of sex-specific regulation remains unclear. We hypothesized that Sf1 is repressed by the transcription factor Forkhead box L2 (FOXL2) during ovarian development. In an in vitro system (TM3 cells), up-regulation of Sf1 by the WT1 splice variant WT1-KTS was antagonized by FOXL2, as determined by quantitative RT-PCR. Using reporter assays, we localized the Sf1 proximal promoter region involved in this antagonism to a 674-bp interval. A conserved FOXL2 binding site was identified in this interval by in vitro chromatin immunoprecipitation. Introducing mutations into this site abolished negative regulation by FOXL2 in reporter assays. Finally, in Foxl2-null mice, Sf1 expression was increased 2-fold relative to wild-type XX fetal gonads. Our results support the hypothesis that FOXL2 negatively regulates Sf1 expression by antagonizing WT1-KTS during early ovarian development in mice.Takasawa, K., Kashimada, K., Pelosi, E., Takagi, M., Morio, T., Asahara, H., Schlessinger, D., Mizutani, S., Koopman, P. FOXL2 transcriptionally represses Sf1 expression by antagonizing WT1 during ovarian development in mice.