Small ubiquitin-like modifier 1 (SUMO-1) modification of the synergy control motif of Ad4 binding protein/steroidogenic factor 1 (Ad4BP/SF-1) regulates synergistic transcription between Ad4BP/SF-1 and Sox9
Small ubiquitin-like modifier 1 (SUMO-1) modification of the synergy control motif of Ad4 binding protein/steroidogenic factor 1 (Ad4BP/SF-1) regulates synergistic transcription between Ad4BP/SF-1 and Sox9
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DOI：
10.1210/me.2004-0173
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发表时间：
2004-10-01
影响因子：
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通讯作者：
Morohashi, KI
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文献类型：
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作者：
Komatsu, T;Mizusaki, H;Morohashi, KI
Ad4结合蛋白/甾体生成因子1 （Ad4BP/SF- 1）是一种孤儿核受体，对甾体生成组织的发育和功能至关重要。为了研究Ad4BP/SF- 1的转录调控，我们进行了双杂交筛选，并分离了sumoylation[偶联一个小泛素样修饰物（SUMO-1）]组分Ubc9、活化STAT 1蛋白抑制剂（PIAS1）和活化STAT 3蛋白抑制剂（PIAS3）。细胞培养和体外研究表明，Ad4BP/SF- 1在K119和K194位点被介导。由于K194位于协同控制（SC）基序中，该基序被定义为抑制含有多个结合位点的启动子的协同转录，因此研究了SC基序的功能与sumoylation之间的相关性。Ad4BP/SF- 1的K194R突变体，不能被summoylation，显示出包含多个Ad4/SF-1位点的启动子的协同转录增强，这表明summoylation是通过SC基序抑制转录协同的必要条件。已经证实，缪勒管抑制物质基因的转录主要受Ad4BP/ SF-1的控制，其转录受Sox9、Gata4和Wt1的协同调节。有趣的是，我们发现所有这些因子都被酰化了，而且这些酰化位点发生在SC基序中。基于对Ad4BP/ SF-1和Sox9的SC基序突变导致其协同转录增强的观察，我们认为SC基序调控了不同类型转录因子之间的协同转录。考虑到这两种突变体都不能被sumo化，sumo化很可能与这种调节有关。由于体外sumoylation Ad4BP/ SF-1发现DNA结合活性和与Sox9的相互作用不受影响，sumo酰化可能通过影响组成转录复合物的因子之间的选择性和合作相互作用来调节转录。
An orphan nuclear receptor, Ad4 binding protein/ steroidogenic factor 1 (Ad4BP/SF- 1), is essential for the development and function of steroidogenic tissues. To examine the transcriptional regulation of Ad4BP/SF- 1, two-hybrid screening was performed, and the sumoylation [ conjugation of a small ubiqutin-like modifier (SUMO-1)] components Ubc9, protein inhibitor of activated STAT 1 (PIAS1), and protein inhibitor of activated STAT 3 (PIAS3) were isolated. Cultured cell and in vitro studies revealed that Ad4BP/SF- 1 is sumoylated at K119 and K194. Because K194 lies within the synergy control (SC) motif defined to repress synergistic transcription from promoters containing multiple binding sites, correlation between the functions of the SC motif and sumoylation was investigated. The K194R mutant of Ad4BP/SF- 1, which cannot be sumoylated, showed enhanced synergistic transcription from a promoter containing multiple Ad4/SF-1 sites, suggesting that sumoylation is necessary for repression of transcriptional synergy through the SC motif. It has been established that the Mullerian inhibiting substance gene is transcribed predominantly under the control of Ad4BP/ SF-1 and, moreover, its transcription is regulated synergistically with Sox9, Gata4, and Wt1. Interestingly, it was found that all of these factors are sumoylated, and these sumoylation sites occur within SC motifs. Based on the observation that SC motif mutants of Ad4BP/ SF-1 and Sox9 resulted in the enhancement of their synergistic transcription, it was concluded that the SC motif regulates synergistic transcription even between distinct types of transcription factors. Considering that both mutants cannot be sumoylated, it is likely that sumoylation is implicated in this regulation. Because it was revealed with an in vitro sumoylated Ad4BP/ SF-1 that DNA binding activity and interaction with Sox9 were unaffected, sumoylation may regulate transcription through affecting selective and cooperative interaction among factors constituting transcriptional complexes.