Organellar Glue: A Molecular Tool to Artificially Control Chloroplast–Chloroplast Interactions

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.

[1]  Y. Kodama,et al.  Chloroplast relocation movement in the liverwort Apopellia endiviifolia , 2021, Physiologia plantarum.

[2]  K. Numata,et al.  Mitochondrial movement during its association with chloroplasts in Arabidopsis thaliana , 2021, Communications biology.

[3]  Florian A. Busch,et al.  Photorespiration in the context of Rubisco biochemistry, CO2 diffusion, and metabolism. , 2020, The Plant journal : for cell and molecular biology.

[4]  M. Hayashi,et al.  Re-evaluation of physical interaction between plant peroxisomes and other organelles using live-cell imaging techniques. , 2019, Journal of integrative plant biology.

[5]  Y. Kodama,et al.  A novel orange-colored bimolecular fluorescence complementation (BiFC) assay using monomeric Kusabira-Orange protein. , 2018, BioTechniques.

[6]  Y. Kodama,et al.  Particle bombardment and subcellular protein localization analysis in the aquatic plant Egeria densa , 2017, PeerJ.

[7]  A. Holzinger,et al.  Chloroplast aggregation during the cold-positioning response in the liverwort Marchantia polymorpha , 2017, Journal of Plant Research.

[8]  Y. Kodama Time Gating of Chloroplast Autofluorescence Allows Clearer Fluorescence Imaging In Planta , 2016, PloS one.

[9]  Tessa M. Burch-Smith,et al.  Chloroplast signaling within, between and beyond cells , 2015, Front. Plant Sci..

[10]  Yoichiroh Hosokawa,et al.  Physical interaction between peroxisomes and chloroplasts elucidated by in situ laser analysis , 2015, Nature Plants.

[11]  Yutaka Kodama,et al.  Bimolecular fluorescence complementation (BiFC): a 5-year update and future perspectives. , 2012, BioTechniques.

[12]  D. Thirumalai,et al.  Denaturant-dependent folding of GFP , 2012, Proceedings of the National Academy of Sciences.

[13]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[14]  T. Kuromori,et al.  SD3, an Arabidopsis thaliana homolog of TIM21, affects intracellular ATP levels and seedling development. , 2012, Molecular plant.

[15]  H. Fukada,et al.  Structure and characteristics of reassembled fluorescent protein, a new insight into the reassembly mechanisms. , 2011, Bioorganic & medicinal chemistry letters.

[16]  Yutaka Kodama,et al.  An improved bimolecular fluorescence complementation assay with a high signal-to-noise ratio. , 2010, BioTechniques.

[17]  Y. Kodama,et al.  Low temperature-induced chloroplast relocation mediated by a blue light receptor, phototropin 2, in fern gametophytes , 2008, Journal of Plant Research.

[18]  A. Tanaka Photosynthetic activity in winter needles of the evergreen tree Taxus cuspidata at low temperatures. , 2007, Tree physiology.

[19]  Han Liu,et al.  Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis under physiological conditions. , 2006, BioTechniques.

[20]  Y. Lee,et al.  Identification of a Signal That Distinguishes between the Chloroplast Outer Envelope Membrane and the Endomembrane System in Vivo , 2001, The Plant Cell Online.

[21]  C. Siegerist,et al.  Reproducible Imaging and Dissection of Plasmid DNA Under Liquid with the Atomic Force Microscope , 1992, Science.

[22]  William D. Richardson,et al.  A short amino acid sequence able to specify nuclear location , 1984, Cell.