Department of Chemistry, Central Washington University, 400 E. University Way, Ellensburg, WA, 98929-7539, USA.
Division of Chemistry and Chemical Engineering, California Institute of Technology, 157 Broad Center, M/C, Pasadena, USA.
Protein J. 2019 Feb;38(1):2-11. doi: 10.1007/s10930-019-09810-1.
The mammalian homolog of Drosophila empty spiracles 2 (Emx2) is a homeobox transcription factor that plays central roles in early development of the inner ear, pelvic and shoulder girdles, cerebral cortex, and urogenital organs. The role for Emx2 is best understood within the context of the development of the neocortical region of the cortex, where Emx2 is expressed in a high posterior-medial to low anterior-lateral gradient that regulates the partitioning of the neocortex into different functional fields that perform discrete computational tasks. Despite several lines of evidence demonstrating an Emx2 concentration-dependent mechanism for establishing functional areas within the developing neocortex, little is known about how Emx2 physically carries out this role. Although several binding partners for Emx2 have been identified (including Sp8, eIF4E, and Pbx1), no screens have been used to identify potential protein binding partners for this protein. We utilized a yeast two-hybrid screen using a library constructed from embryonic mouse cDNA in an attempt to identify novel binding partners for Emx2. This initial screen isolated two potential Emx2-binding partner proteins, Cnot6l and QkI-7. These novel Emx2-binding proteins are involved in multiple levels of mRNA metabolism that including splicing, mRNA export, translation, and destruction, thus making them interesting targets for further study.
果蝇空泡基因 2(Emx2)的哺乳动物同源物是一种同源盒转录因子,在耳朵内、骨盆和肩带、大脑皮层和泌尿生殖器官的早期发育中发挥核心作用。Emx2 的作用在皮层新皮层区域的发育背景下最容易理解,在该区域,Emx2 以高后-中至高前-低的梯度表达,调节新皮层分为执行离散计算任务的不同功能区。尽管有几条证据表明 Emx2 存在浓度依赖性机制,可在发育中的新皮层内建立功能区,但对于 Emx2 如何发挥这种作用知之甚少。尽管已经确定了几个 Emx2 的结合伴侣(包括 Sp8、eIF4E 和 Pbx1),但尚未使用筛选方法来鉴定该蛋白的潜在蛋白结合伴侣。我们利用酵母双杂交筛选,使用从胚胎期小鼠 cDNA 构建的文库,试图鉴定 Emx2 的新结合伴侣蛋白。该初始筛选分离出两个潜在的 Emx2 结合伴侣蛋白,Cnot6l 和 QkI-7。这些新的 Emx2 结合蛋白参与多个水平的 mRNA 代谢,包括剪接、mRNA 输出、翻译和破坏,因此成为进一步研究的有趣目标。