Fujimoto Yukari, Fujimoto Masaya, Konno Daijiro, Nakazawa Naotaka
Graduate School of Science and Engineering, Kindai University, Higashiosaka-city, Osaka, Japan.
Faculty of Science and Engineering, Kindai University, Higashiosaka-city, Osaka, Japan.
MicroPubl Biol. 2025 Aug 28;2025. doi: 10.17912/micropub.biology.001669. eCollection 2025.
Four-and-a-half LIM domains 2 (FHL2) is a molecule that plays a key role in cell proliferation in response to mechanical signals. It shuttles between focal adhesions (FAs) or stress fibers (SFs) and the nucleus in a force-dependent manner. FHL2 interacts with other cytoskeletal molecules at FAs and SFs, but FHL2 interacts with transcriptional factors in the nucleus. This leads to modulation of gene expression for cell proliferation. However, the overall picture of interacting proteins with FHL2 at different regions is poorly understood. Here, we report a stable cell line that expresses FHL2-GFP-BirA, a fusion protein of FHL2 with biotin ligase. FHL2-GFP-BirA localizes at the FAs but accumulates in the nucleus after myosin II inhibition, exhibiting behavior similar to endogenous FHL2. These results suggest that the BioID technique can be used to identify proteins interacting with FHL2 under different mechanical conditions.
四又二分之一LIM结构域蛋白2（FHL2）是一种在细胞对机械信号的增殖反应中起关键作用的分子。它以力依赖的方式在粘着斑（FAs）或应力纤维（SFs）与细胞核之间穿梭。FHL2在粘着斑和应力纤维处与其他细胞骨架分子相互作用，但在细胞核中与转录因子相互作用。这导致细胞增殖相关基因表达的调控。然而，对于不同区域与FHL2相互作用的蛋白质的整体情况了解甚少。在此，我们报道了一种稳定表达FHL2-GFP-BirA的细胞系，FHL2-GFP-BirA是FHL2与生物素连接酶的融合蛋白。FHL2-GFP-BirA定位于粘着斑，但在肌球蛋白II抑制后在细胞核中积累，表现出与内源性FHL2相似的行为。这些结果表明，BioID技术可用于鉴定在不同机械条件下与FHL2相互作用的蛋白质。
