Department of Neurobiology, School of Biological Sciences, University of California, San Diego, San Diego, CA 92093, USA.
Department of Cell and Developmental Biology, School of Biological Sciences, University of California, San Diego, San Diego, CA 92093, USA.
Cell Rep. 2024 Oct 22;43(10):114776. doi: 10.1016/j.celrep.2024.114776. Epub 2024 Sep 20.
The EFA6 protein family, originally identified as Sec7 guanine nucleotide exchange factors, has also been found to regulate cortical microtubule (MT) dynamics. Here, we find that in the mature C. elegans epidermal epithelium, EFA-6 forms punctate foci in specific regions of the apical cortex, dependent on its intrinsically disordered region (IDR). The EFA-6 IDR can form biomolecular condensates in vitro. In genetic screens for mutants with altered GFP::EFA-6 localization, we identified a gain-of-function (gf) mutation in α-tubulin tba-1 that induces ectopic EFA-6 foci in multiple cell types. Lethality of tba-1(gf) is partially suppressed by loss of function in efa-6. The ability of TBA-1(gf) to trigger ectopic EFA-6 foci requires β-tubulin TBB-2 and the chaperon EVL-20/Arl2. tba-1(gf)-induced EFA-6 foci display slower turnover, contain the MT-associated protein TAC-1/TACC, and require the EFA-6 MT elimination domain (MTED). Our results reveal functionally important crosstalk between cellular tubulins and cortical MT regulators in vivo.
EFA6 蛋白家族最初被鉴定为 Sec7 鸟嘌呤核苷酸交换因子,也被发现可调节皮质微管 (MT) 动力学。在这里,我们发现,在成熟的 C. elegans 表皮上皮中,EFA-6 在顶皮质的特定区域形成点状焦点,这依赖于其无规卷曲区域 (IDR)。EFA-6 IDR 可以在体外形成生物分子凝聚物。在 GFP::EFA-6 定位改变的突变体的遗传筛选中,我们在α-微管蛋白 tba-1 中发现了一个功能获得 (gf) 突变,该突变导致多个细胞类型中 EFA-6 焦点异位形成。tba-1(gf)的致死性部分被 efa-6 的功能丧失所抑制。TBA-1(gf) 触发异位 EFA-6 焦点的能力需要β-微管蛋白 TBB-2 和伴侣蛋白 EVL-20/Arl2。tba-1(gf)诱导的 EFA-6 焦点显示出较慢的周转率,包含 MT 相关蛋白 TAC-1/TACC,并需要 EFA-6 MT 消除结构域 (MTED)。我们的结果揭示了细胞微管蛋白和皮质 MT 调节剂之间在体内具有功能上重要的相互作用。