Center for Bioscience Research and Education, Utsunomiya University, Tochigi 321-8505, Japan.
Graduate School of Regional Development and Creativity, Utsunomiya University, Tochigi 321-8505, Japan.
ACS Synth Biol. 2022 Oct 21;11(10):3190-3197. doi: 10.1021/acssynbio.2c00367. Epub 2022 Sep 30.
Organelles can physically interact to facilitate various cellular processes such as metabolite exchange. Artificially regulating these interactions represents a promising approach for synthetic biology. Here, we artificially controlled chloroplast-chloroplast interactions in living plant cells with our organelle glue (ORGL) technique, which is based on reconstitution of a split fluorescent protein. We simultaneously targeted N-terminal and C-terminal fragments of a fluorescent protein to the chloroplast outer envelope membrane or cytosol, respectively, which induced chloroplast-chloroplast interactions. The cytosolic C-terminal fragment likely functions as a bridge between two N-terminal fragments, thereby bringing the chloroplasts in close proximity to interact. We modulated the frequency of chloroplast-chloroplast interactions by altering the ratio of N- and C-terminal fragments. We conclude that the ORGL technique can successfully control chloroplast-chloroplast interactions in plants, providing a proof of concept for the artificial regulation of organelle interactions in living cells.
细胞器可以通过物理相互作用来促进各种细胞过程，例如代谢物交换。人为调节这些相互作用是合成生物学的一种有前途的方法。在这里，我们使用基于荧光蛋白分裂重组成分的细胞器胶（ORGL）技术，在活植物细胞中人为控制叶绿体-叶绿体相互作用。我们分别将荧光蛋白的 N 端和 C 端片段靶向叶绿体的外膜或细胞质，从而诱导叶绿体-叶绿体相互作用。细胞质 C 端片段可能作为两个 N 端片段之间的桥梁，从而使叶绿体紧密相互作用。我们通过改变 N 端和 C 端片段的比例来调节叶绿体-叶绿体相互作用的频率。我们得出结论，ORGL 技术可以成功地在植物中控制叶绿体-叶绿体相互作用，为在活细胞中人工调节细胞器相互作用提供了概念验证。
