Kiefer Susan McLeskey, McDill Bradley W, Yang Jing, Rauchman Michael
Renal Division, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
J Biol Chem. 2002 Apr 26;277(17):14869-76. doi: 10.1074/jbc.M200052200. Epub 2002 Feb 8.
The multi-zinc finger proteins of the Sal family regulate organogenesis. Genetic evidence from Drosophila has shown that spalt (sal) can alter gene expression in a cell autonomous fashion, but Sal proteins have never been directly analyzed for their ability to activate or repress transcription. In this report, we show that a member of the Sal family, mouse Sall1, is a potent transcriptional repressor. When fused to a heterologous DNA-binding domain, Sall1 represses transcription of a luciferase reporter by over 100-fold. Expression of the N terminus alone is sufficient for dose-responsive repression that, as shown by deletion analysis, requires the extreme N-terminal amino acids of the protein. The N terminus of Sall1 can repress at both short and long range relative to the promoter, and treatment with the histone deacetylase (HDAC) inhibitor, trichostatin A, alleviates repression by 3-fold. The same regions of the protein that are required for repression physically interact with components of chromatin remodeling complexes, HDAC1, HDAC2, RbAp46/48, MTA-1, and MTA-2. Finally, we demonstrate that Sall1 is localized to discrete nuclear foci and this localization depends on the N-terminal repression domain. Together, these results suggest that the N terminus of mouse Sall1 can recruit HDAC complexes to mediate transcriptional repression.
Sal家族的多锌指蛋白调控器官发生。来自果蝇的遗传学证据表明,spalt(sal)能够以细胞自主的方式改变基因表达,但从未直接分析过Sal蛋白激活或抑制转录的能力。在本报告中,我们表明Sal家族的一个成员,小鼠Sall1,是一种有效的转录抑制因子。当与异源DNA结合结构域融合时,Sall1可将荧光素酶报告基因的转录抑制100倍以上。单独表达N端就足以实现剂量依赖性抑制,缺失分析表明,这需要该蛋白极端的N端氨基酸。相对于启动子,Sall1的N端在短距离和长距离都能发挥抑制作用,用组蛋白去乙酰化酶(HDAC)抑制剂曲古抑菌素A处理可使抑制作用减轻3倍。该蛋白抑制作用所需的相同区域与染色质重塑复合物的组分HDAC1、HDAC2、RbAp46/48、MTA-1和MTA-2发生物理相互作用。最后,我们证明Sall1定位于离散的核灶,且这种定位依赖于N端抑制结构域。总之,这些结果表明小鼠Sall1的N端可募集HDAC复合物来介导转录抑制。