Montreal Clinical Research Institute (IRCM), Montréal, QC H2W 1R7, Canada.
Molecular Biology Programs, Université de Montréal, Montréal, QC H3T 1J4, Canada.
J Cell Sci. 2024 May 1;137(9). doi: 10.1242/jcs.262140. Epub 2024 May 14.
The ADP-ribosylation factors (ARFs) and ARF-like (ARL) GTPases serve as essential molecular switches governing a wide array of cellular processes. In this study, we used proximity-dependent biotin identification (BioID) to comprehensively map the interactome of 28 out of 29 ARF and ARL proteins in two cellular models. Through this approach, we identified ∼3000 high-confidence proximal interactors, enabling us to assign subcellular localizations to the family members. Notably, we uncovered previously undefined localizations for ARL4D and ARL10. Clustering analyses further exposed the distinctiveness of the interactors identified with these two GTPases. We also reveal that the expression of the understudied member ARL14 is confined to the stomach and intestines. We identified phospholipase D1 (PLD1) and the ESCPE-1 complex, more precisely, SNX1, as proximity interactors. Functional assays demonstrated that ARL14 can activate PLD1 in cellulo and is involved in cargo trafficking via the ESCPE-1 complex. Overall, the BioID data generated in this study provide a valuable resource for dissecting the complexities of ARF and ARL spatial organization and signaling.
ADP-核糖基化因子 (ARF) 和 ARF 样 (ARL) GTPases 作为重要的分子开关，调节着广泛的细胞过程。在这项研究中，我们使用邻近依赖性生物素鉴定 (BioID) 技术，在两种细胞模型中全面绘制了 29 个 ARF 和 ARL 蛋白中的 28 个的互作组。通过这种方法，我们鉴定了约 3000 个高可信度的近端相互作用蛋白，使我们能够对家族成员进行亚细胞定位。值得注意的是，我们揭示了 ARL4D 和 ARL10 的先前未定义的定位。聚类分析进一步揭示了这两种 GTPase 所鉴定的相互作用蛋白的独特性。我们还揭示了研究较少的 ARL14 成员的表达局限于胃和肠道。我们鉴定了磷脂酶 D1 (PLD1) 和 ESCPE-1 复合物，更确切地说是 SNX1，作为邻近相互作用蛋白。功能测定表明，ARL14 可以在细胞内激活 PLD1，并通过 ESCPE-1 复合物参与货物运输。总体而言，本研究中生成的 BioID 数据为解析 ARF 和 ARL 空间组织和信号转导的复杂性提供了有价值的资源。
