School of Life and Environmental Sciences, University of Sydney, NSW, 2006, Australia.
Teva Pharmaceuticals Australia Pty Ltd, Macquarie Park, NSW, 2113, Australia.
Sci Rep. 2017 Jul 4;7(1):4579. doi: 10.1038/s41598-017-04587-8.
LIM-Homeodomain (LIM-HD) transcription factors are highly conserved in animals where they are thought to act in a transcriptional 'LIM code' that specifies cell types, particularly in the central nervous system. In chick and mammals the interaction between two LIM-HD proteins, LHX3 and Islet1 (ISL1), is essential for the development of motor neurons. Using yeast two-hybrid analysis we showed that the Caenorhabditis elegans orthologs of LHX3 and ISL1, CEH-14 and LIM-7 can physically interact. Structural characterisation of a complex comprising the LIM domains from CEH-14 and a LIM-interaction domain from LIM-7 showed that these nematode proteins assemble to form a structure that closely resembles that of their vertebrate counterparts. However, mutagenic analysis across the interface indicates some differences in the mechanisms of binding. We also demonstrate, using fluorescent reporter constructs, that the two C. elegans proteins are co-expressed in a small subset of neurons. These data show that the propensity for LHX3 and Islet proteins to interact is conserved from C. elegans to mammals, raising the possibility that orthologous cell specific LIM-HD-containing transcription factor complexes play similar roles in the development of neuronal cells across diverse species.
LIM 同源结构域(LIM-HD)转录因子在动物中高度保守,据认为它们在转录“LIM 密码”中起作用,特别是在中枢神经系统中,决定细胞类型。在鸡和哺乳动物中,两个 LIM-HD 蛋白 LHX3 和 Islet1(ISL1)之间的相互作用对于运动神经元的发育是必不可少的。通过酵母双杂交分析,我们表明 LHX3 和 ISL1 的秀丽隐杆线虫同源物 CEH-14 和 LIM-7 可以物理相互作用。包含 CEH-14 的 LIM 结构域和 LIM-7 的 LIM 相互作用结构域的复合物的结构特征表明,这些线虫蛋白组装形成的结构与它们的脊椎动物对应物非常相似。然而,跨界面的诱变分析表明,结合的机制存在一些差异。我们还使用荧光报告构建体证明,两种秀丽隐杆线虫蛋白在一小部分神经元中共同表达。这些数据表明,LHX3 和 Islet 蛋白相互作用的倾向从秀丽隐杆线虫到哺乳动物是保守的,这增加了这样一种可能性,即同源细胞特异性 LIM-HD 包含转录因子复合物在不同物种的神经元细胞发育中发挥类似的作用。