The Dilute domain of Canoe is not essential for Canoe's role in linking adherens junctions to the cytoskeleton but contributes to robustness of morphogenesis.
The Dilute domain of Canoe is not essential for Canoe's role in linking adherens junctions to the cytoskeleton but contributes to robustness of morphogenesis.
复制标题
Canoe 的稀释结构域对于 Canoe 连接粘附连接与细胞骨架的作用不是必需的，但有助于形态发生的稳健性。
DOI：
10.1101/2023.10.18.562854
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Peifer,Mark
中科院分区：
文献类型：
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作者：
McParland,EmilyD;AmberButcher,T;Gurley,NoahJ;Johnson,RuthI;Slep,KevinC;Peifer,Mark
细胞-细胞之间牢固的连接连接和肌动蛋白细胞骨架允许细胞在形态发生过程中改变形状和移动，而不会撕裂组织。果蝇CANOE及其哺乳动物同源基因Afadin的多结构域蛋白对这种联系至关重要，在它们缺失的情况下，许多形态发生事件都失败了。为了定义潜在的机制，我们正在拆解独木舟，使用果蝇作为我们的模型。独木舟和Afadin共享五个折叠的蛋白质结构域，紧随其后的是一个巨大的内在无序区域。这些折叠结构域中最大的是稀释域，它存在于Canoe/Afadin、它们的近亲以及MyosinV家族成员中。为了确定独木舟稀释域的作用，我们结合了生化、遗传和细胞生物学分析。使用AlphaFold工具揭示了预测的Canoe/Afadin稀释域的结构，提供了与MyosinV的相似和对比。我们的生化数据表明了一个潜在的共同功能：二聚体的能力。接下来，我们产生了果蝇突变体，其稀释域被彻底删除。令人惊讶的是，这些突变体是活的和可育的，并且独木舟ΔDil蛋白定位于附着的连接处，并在张力下的连接处富含。然而，当我们在致敏试验中减少CanoeΔDil蛋白的剂量时，很明显它不能提供全部的野生型功能。此外，独木舟Δdil突变体在蛹眼发育方面存在缺陷，这是另一个需要精心安排细胞重排的过程。总之，这些数据揭示了AJ-细胞骨架连接在多个胚胎和胚胎后事件中的稳健性，以及自然选择即使在强大的系统中也能维持蛋白质结构的能力。
Robust linkage between cell-cell adherens junctions and the actomyosin cytoskeleton allows cells to change shape and move during morphogenesis without tearing tissues apart. The multidomain protein Drosophila Canoe and its mammalian homolog Afadin are critical for this linkage, and in their absence many events of morphogenesis fail. To define underlying mechanisms, we are taking Canoe apart, using Drosophila as our model. Canoe and Afadin share five folded protein domains, followed by a large intrinsically disordered region. The largest of these folded domains is the Dilute domain, which is found in Canoe/Afadin, their paralogs, and members of the MyosinV family. To define the roles of Canoe’s Dilute domain we have combined biochemical, genetic and cell biological assays. Use of the AlphaFold tools revealed the predicted structure of the Canoe/Afadin Dilute domain, providing similarities and contrasts with that of MyosinV. Our biochemical data suggest one potential shared function: the ability to dimerize. We next generated Drosophila mutants with the Dilute domain cleanly deleted. Surprisingly, these mutants are viable and fertile, and CanoeΔDIL protein localizes to adherens junctions and is enriched at junctions under tension. However, when we reduce the dose of CanoeΔDIL protein in a sensitized assay, it becomes clear it does not provide full wildtype function. Further, canoeΔDIL mutants have defects in pupal eye development, another process that requires orchestrated cell rearrangements. Together, these data reveal the robustness in AJ-cytoskeletal connections during multiple embryonic and postembryonic events, and the power of natural selection to maintain protein structure even in robust systems.