Sasahara Miyu, Matsumoto Shunsuke, Tamura Yasushi
Graduate School of Science and Engineering, Yamagata University.
Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University.
MicroPubl Biol. 2025 Aug 19;2025. doi: 10.17912/micropub.biology.001780. eCollection 2025.
In yeast, mitochondrial fission is mediated by the dynamin-like GTPase Dnm1, which is recruited to the mitochondrial outer membrane by its receptor, Fis1. To investigate the spatial distribution of Fis1, we used the CRISPR-Cas9 system to insert the gene fragment encoding mNeonGreen into the gene for its N-terminal tagging. Fluorescence microscopy revealed that mNeonGreen-Fis1 appeared as discrete puncta on mitochondria, in addition to a diffuse signal. Here, we show that the focal clustering of Fis1 is dependent on Dnm1. Our findings provide insight into the spatial organization of membrane proteins, highlighting a mechanism by which a downstream effector can influence the distribution of its upstream receptor.
在酵母中，线粒体分裂由动力蛋白样GTP酶Dnm1介导，Dnm1通过其受体Fis1被招募到线粒体外膜。为了研究Fis1的空间分布，我们使用CRISPR-Cas9系统将编码mNeonGreen的基因片段插入到其基因中进行N端标记。荧光显微镜显示，除了弥散信号外，mNeonGreen-Fis1在线粒体上表现为离散的斑点。在这里，我们表明Fis1的局灶性聚集依赖于Dnm1。我们的发现为膜蛋白的空间组织提供了见解，突出了一种下游效应器可以影响其上游受体分布的机制。
