A network of interacting transcriptional regulators involved in Drosophila neural fate specification revealed by the yeast two-hybrid system
A network of interacting transcriptional regulators involved in Drosophila neural fate specification revealed by the yeast two-hybrid system
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DOI：
10.1073/pnas.94.24.13099
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发表时间：
1997-11-25
影响因子：
11.1
通讯作者：
Delidakis, C
中科院分区：
文献类型：
--
作者：
Alifragis, P;Poortinga, G;Delidakis, C
果蝇中的神经命运特化由前神经基因的产物促进，例如无毛-盾壳复合体的产物，并由分裂增强子[E（spl）]复合体、毛状体和extramacrochaetae的产物拮抗。由于所有这些蛋白都具有螺旋-环螺旋（HLH）二聚化结构域，我们使用酵母双杂交系统研究了它们潜在的成对相互作用。该系统的保真度建立了密切再现Da，Ac，Sc和Extramacrochaetae之间的相互作用已经记录的能力。我们发现，七个E（SPL）的基本HLH蛋白可以形成家庭和异源二聚体之间的不同偏好。我们进一步表明，E（spl）蛋白的一个子集可以与Da异源二聚化，另一个子集可以与前神经蛋白异源二聚化，还有一个子集可以与两者都异源二聚化，表明E（spl）家族内的特化。Hairy不显示与任何测试的HLH蛋白的相互作用，它确实与非HLH蛋白Groucho相互作用，Groucho本身与所有E（spl）碱性HLH蛋白相互作用，但没有原神经蛋白或DA。我们研究了这些相互作用的一些位点特异性和缺失突变的结构要求。
Neural fate specification in Drosophila is promoted by the products of the proneural genes, such as those of the achaete-scute complex, and antagonized by the products of the Enhancer of split [E(spl)] complex, hairy, and extramacrochaetae. As all these proteins bear a helix-loop helix (HLH) dimerization domain, we investigated their potential pairwise interactions using the yeast two hybrid system. The fidelity of the system was established by its ability to closely reproduce the already documented interactions among Da, Ac, Sc, and Extramacrochaetae. We show that the seven E(spl) basic HLH proteins can form home-and heterodimers inter-se with distinct preferences. We further show that a subset of E(spl) proteins can heterodimerize with Da, another subset can heterodimerize with proneural proteins, and yet another with both, indicating specialization within the E(spl) family, Hairy displays no interactions with any of the HLH proteins tested, It does interact with the non-HLH protein Groucho, which itself interacts with all E(spl) basic HLH proteins, but with none of the proneural proteins or Da. We investigated the structural requirements for some of these interactions by site-specific and deletion mutagenesis.