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微管调节蛋白EFA-6形成空间受限的皮质病灶,这依赖于其内在无序区域以及与微管蛋白的相互作用。

The microtubule regulator EFA-6 forms spatially restricted cortical foci dependent on its intrinsically disordered region and interactions with tubulins.

作者信息

Sandhu Anjali, Lyu Xiaohui, Wan Xinghaoyun, Meng Xuefeng, Tang Ngang Heok, Gonzalez Gilberto, Syed Ishana N, Chen Lizhen, Jin Yishi, Chisholm Andrew D

机构信息

Department of Neurobiology, School of Biological Sciences, University of California San Diego, CA 92093 USA.

Department of Cell and Developmental Biology, School of Biological Sciences, University of California San Diego, CA 92093 USA.

出版信息

bioRxiv. 2024 Apr 14:2024.04.14.588158. doi: 10.1101/2024.04.14.588158.

Abstract

Microtubules (MTs) are dynamic components of the cytoskeleton and play essential roles in morphogenesis and maintenance of tissue and cell integrity. Despite recent advances in understanding MT ultrastructure, organization, and growth control, how cells regulate MT organization at the cell cortex remains poorly understood. The EFA-6/EFA6 proteins are recently identified membrane-associated proteins that inhibit cortical MT dynamics. Here, combining visualization of endogenously tagged EFA-6 with genetic screening, we uncovered tubulin-dependent regulation of EFA-6 patterning. In the mature epidermal epithelium, EFA-6 forms punctate foci in specific regions of the apical cortex, dependent on its intrinsically disordered region (IDR). We further show the EFA-6 IDR is sufficient to form biomolecular condensates . In screens for mutants with altered GFP::EFA-6 localization, we identified a novel gain-of-function (gf) mutation in an α-tubulin that induces ectopic EFA-6 foci in multiple cell types. animals exhibit temperature-sensitive embryonic lethality, which is partially suppressed by , indicating the interaction between tubulins and EFA-6 is important for normal development. TBA-1(gf) shows reduced incorporation into filamentous MTs but has otherwise mild effects on cellular MT organization. The ability of TBA-1(gf) to trigger ectopic EFA-6 foci formation requires β-tubulin TBB-2 and the chaperon EVL-20/Arl2. The induced EFA-6 foci display slower turnover, contain the MT-associated protein TAC-1/TACC, and require the EFA-6 MTED. Our results reveal a novel crosstalk between cellular tubulins and cortical MT regulators .

摘要

微管(MTs)是细胞骨架的动态组成部分,在组织形态发生以及组织和细胞完整性的维持中发挥着重要作用。尽管在理解微管超微结构、组织和生长控制方面取得了最新进展,但细胞如何在细胞皮层调节微管组织仍知之甚少。EFA-6/EFA6蛋白是最近鉴定出的抑制皮层微管动态的膜相关蛋白。在这里,我们将内源性标记的EFA-6可视化与基因筛选相结合,发现了EFA-6模式的微管蛋白依赖性调节。在成熟的表皮上皮中,EFA-6在顶端皮层的特定区域形成点状病灶,这取决于其内在无序区域(IDR)。我们进一步表明,EFA-6的IDR足以形成生物分子凝聚物。在筛选绿色荧光蛋白::EFA-6定位改变的突变体时,我们在α-微管蛋白中鉴定出一种新的功能获得(gf)突变,该突变在多种细胞类型中诱导异位EFA-6病灶。 动物表现出温度敏感的胚胎致死性, 可部分抑制这种致死性,这表明微管蛋白与EFA-6之间的相互作用对正常发育很重要。TBA-1(gf)显示出较少掺入丝状微管,但对细胞微管组织的其他影响较小。TBA-1(gf)触发异位EFA-6病灶形成的能力需要β-微管蛋白TBB-2和伴侣蛋白EVL-20/Arl2。诱导的EFA-6病灶显示出较慢的周转,包含微管相关蛋白TAC-1/TACC,并且需要EFA-6的微管末端结构域(MTED)。我们的结果揭示了细胞微管蛋白与皮层微管调节剂之间的一种新的相互作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ef5/11030407/5bd697857278/nihpp-2024.04.14.588158v1-f0001.jpg

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