A functional interaction between the histone deacetylase Rpd3 and the corepressor Groucho in Drosophila development
A functional interaction between the histone deacetylase Rpd3 and the corepressor Groucho in Drosophila development
复制标题
DOI：
10.1101/gad.13.17.2218
复制
发表时间：
1999-09-01
影响因子：
10.5
通讯作者：
Courey, AJ
中科院分区：
文献类型：
--
作者：
Chen, GQ;Fernandez, J;Courey, AJ
果蝇的groucho（gro）基因编码一种转录共抑制因子，它在许多发育过程中起着关键作用。为了阐明Gro介导的抑制机制，我们利用Gro作为亲和试剂从胚胎核提取物中纯化Gro结合蛋白。其中一种蛋白被发现是组蛋白去乙酰化酶Rpd3。蛋白质 - 蛋白质相互作用实验表明，Gro和Rpd3在体内形成复合物，并且它们通过Gro中富含甘氨酸/脯氨酸（GP）的结构域直接相互作用。细胞培养实验表明，Rpd3增强了GP结构域的抑制作用。此外，使用组蛋白去乙酰化酶抑制剂以及无催化活性的Rpd3形式的实验表明，组蛋白去乙酰化酶活性是高效的Gro介导的抑制所必需的。最后，gro和rpd3的突变对胚胎致死率和模式形成具有协同效应。这些发现支持这样一种观点，即Gro至少部分通过将组蛋白去乙酰化酶Rpd3直接招募到模板上来介导抑制，在模板处它可以调节局部染色质结构。它们也为Rpd3在早期发育中的特定作用提供了证据。
The Drosophila gene groucho (gro) encodes a transcriptional corepressor that has critical roles in many development processes. In an effort to illuminate the mechanism of Gro-mediated repression, we have employed Gro as an affinity reagent to purify Gro-binding proteins from embryonic nuclear extracts. One of these proteins was found to be the histone deacetylase Rpd3. Protein-protein interaction assays suggest that Gro and Rpd3 form a complex in vivo and that they interact directly via the glycine/proline rich (GP) domain in Gro. Cell culture assays demonstrate that Rpd3 potentiates repression by the GP domain. Furthermore, experiments employing a histone deacetylase inhibitor, as well as a catalytically inactive form of Rpd3, imply that histone deacetylase activity is required for efficient Gro-mediated repression. Finally, mutations in gro and rpd3 have synergistic effects on embryonic lethality and pattern formation. These findings support the view that Gro mediates repression, at least in part, by the direct recruitment of the histone deacetylase Rpd3 to the template, where it can modulate local chromatin structure. They also provide evidence for a specific role of Rpd3 in early development.