Subcellular dynamics of estrogen-related receptors involved in transrepression through interactions with scaffold attachment factor B1
Subcellular dynamics of estrogen-related receptors involved in transrepression through interactions with scaffold attachment factor B1
复制标题
DOI：
10.1007/s00418-021-01998-7
复制
发表时间：
2021-06-15
影响因子：
2.3
通讯作者：
Tanaka，Masaki
中科院分区：
文献类型：
--
作者：
Tanida，Takashi;Matsuda，Ken Ichi;Tanaka，Masaki
雌激素相关受体（Estrogen-related receptor，ERR）是核受体超家族的成员之一，与雌激素受体有很强的同源性，由α、β、γ三种亚型组成。没有内源性配体已被确定为ERRs，但他们在代谢，激素和发育过程中发挥关键作用的转录因子没有配体结合。虽然亚核动力学是必不可少的核事件，包括核受体介导的转录调控，ERRs的动力学知之甚少。在这里，我们报告说，雌激素受体显示亚细胞动力学变化，在响应己烯雌酚（DES），合成雌激素，抑制所有三个雌激素受体亚型的反式作用，使用活细胞成像与荧光蛋白标记。经DES处理后，所有ERR亚型在细胞核中形成离散簇，ERRγ也显示核输出。光漂白分析后的荧光恢复显示，DES结合的ERRα和ERRβ的核内迁移率显著降低，DES结合的ERRγ的核内迁移率略有降低。DES处理后，共定位的所有ERR亚型与支架附着因子B1（SAFB 1），核基质相关蛋白，观察到的点样亚核簇，这表明与核基质的ERRs的相互作用。一致地，免疫共沉淀分析证实了DES存在下ERRs和SAFB 1之间的相互作用增强。SAFB 1通过ERR反应元件抑制所有ERR亚型的反式活性。这些结果表明，ERRs在细胞核中的配体依赖性簇形成与SAFB 1介导的反式阻遏密切相关。总之，目前的研究结果提供了一个新的理解的病理生理调节ERR/SAFB 1信号通路及其亚细胞动力学。
Estrogen-related receptor (ERR), a member of the nuclear receptor superfamily, consists of three subtypes (α, β, γ) and has strong homology with estrogen receptor. No endogenous ligands have been identified for ERRs, but they play key roles in metabolic, hormonal, and developmental processes as transcription factors without ligand binding. Although subnuclear dynamics are essential for nuclear events including nuclear receptor-mediated transcriptional regulation, the dynamics of ERRs are poorly understood. Here, we report that ERRs show subcellular kinetic changes in response to diethylstilbestrol (DES), a synthetic estrogen that represses the transactivity of all three ERR subtypes, using live-cell imaging with fluorescent protein labeling. Upon DES treatment, all ERR subtypes formed discrete clusters in the nucleus, with ERRγ also displaying nuclear export. Fluorescence recovery after photobleaching analyses revealed significant reductions in the intranuclear mobility of DES-bound ERRα and ERRβ, and a slight reduction in the intranuclear mobility of DES-bound ERRγ. After DES treatment, colocalization of all ERR subtypes with scaffold attachment factor B1 (SAFB1), a nuclear matrix-associated protein, was observed in dot-like subnuclear clusters, suggesting interactions of the ERRs with the nuclear matrix. Consistently, co-immunoprecipitation analyses confirmed enhanced interactions between ERRs and SAFB1 in the presence of DES. SAFB1 was clarified to repress the transactivity of all ERR subtypes through the ERR-response element. These results demonstrate ligand-dependent cluster formation of ERRs in the nucleus that is closely associated with SAFB1-mediated transrepression. Taken together, the present findings provide a new understanding of the pathophysiology regulated by ERR/SAFB1 signaling pathways and their subcellular dynamics.