Institute of Plant Sciences, University of Regensburg, Universitätsstraße 31, 93053, Regensburg, Germany.
Signalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Schänzlestrasse 1, 79104, Freiburg, Germany.
Plant Cell Rep. 2024 Jun 15;43(7):174. doi: 10.1007/s00299-024-03158-2.
Interactor of WOX2, CDC48A, is crucial for early embryo patterning and shoot meristem stem cell initiation, but is not required for WOX2 protein turnover or subcellular localization. During Arabidopsis embryo patterning, the WUSCHEL HOMEOBOX 2 (WOX2) transcription factor is a major regulator of protoderm and shoot stem cell initiation. Loss of WOX2 function results in aberrant protodermal cell divisions and, redundantly with its paralogs WOX1, WOX3, and WOX5, compromised shoot meristem formation. To elucidate the molecular basis for WOX2 function, we searched for protein interactors by IP-MS/MS from WOX2-overexpression roots displaying reprogramming toward shoot-like cell fates. Here, we report that WOX2 directly interacts with the type II AAA ATPase molecular chaperone CELL DIVISION CYCLE 48A (CDC48A). We confirmed this interaction with bimolecular fluorescence complementation and co-immunoprecipitation and found that both proteins co-localize in the nucleus. We show that CDC48A loss of function results in protoderm and shoot meristem stem cell initiation defects similar to WOX2 loss of function. We also provide evidence that CDC48A promotes WOX2 activity independently of proteolysis or the regulation of nuclear localization, common mechanisms of CDC48A function in other processes. Our results point to a new role of CDC48A in potentiating WOX2 function during early embryo patterning.
WOX2 相互作用因子 CDC48A 对早期胚胎模式形成和茎分生组织干细胞起始至关重要,但对于 WOX2 蛋白周转或亚细胞定位不是必需的。在拟南芥胚胎模式形成过程中,WUSCHEL HOMEOBOX 2(WOX2)转录因子是原表皮和茎分生组织干细胞起始的主要调节因子。WOX2 功能丧失会导致原表皮细胞分裂异常,并且与其同源物 WOX1、WOX3 和 WOX5 冗余,导致茎分生组织形成受损。为了阐明 WOX2 功能的分子基础,我们通过在显示向类似于茎的细胞命运重编程的 WOX2 过表达根中进行 IP-MS/MS 筛选寻找蛋白相互作用因子。在这里,我们报告 WOX2 与 II 型 AAA ATP 酶分子伴侣细胞分裂周期 48A(CDC48A)直接相互作用。我们通过双分子荧光互补和共免疫沉淀证实了这种相互作用,并发现两种蛋白质在核中共定位。我们表明,CDC48A 功能丧失会导致原表皮和茎分生组织干细胞起始缺陷,类似于 WOX2 功能丧失。我们还提供了证据表明,CDC48A 独立于蛋白水解或核定位的调节促进 WOX2 活性,这是 CDC48A 在其他过程中发挥作用的常见机制。我们的研究结果表明,CDC48A 在早期胚胎模式形成过程中增强 WOX2 功能方面具有新的作用。