Abstract
Key message
OsJAZ2 protein has a propensity to form condensates, possibly by multivalent interactions, and can be used to construct artificial compartments in plant cells.
Abstract
Eukaryotic cells contain various membraneless organelles, which are compartments consisting of proteinaceous condensates formed by phase separation. Such compartments are attractive for bioengineering and synthetic biology, because they can modify cellular function by the enrichment of molecules of interest and providing an orthogonal reaction system. This study reports that Oryza sativa JAZ2 protein (OsJAZ2) is an atypical jasmonate signalling regulator that can form large condensates in both the nucleus and cytosol of O. sativa cells. TIFY and Jas domains and low-complexity regions contribute to JAZ2 condensation, possibly by multivalent interaction. Fluorescence recovery after photobleaching (FRAP) analysis suggests that JAZ2 condensates form mostly gel-like or solid compartments, but can also be in a liquid-like state. Deletion of the N-terminal region or the TIFY domain of JAZ2 causes an increase in the mobile fraction of JAZ2 condensates, moderately. Moreover, JAZ2 can also form liquid-like condensates when expressed in Nicotiana benthamiana cells. The recombinant JAZ2 fused to the green fluorescent protein (GFP) forms condensate in vitro, suggesting that the intermolecular interaction of JAZ2 molecules is a driving force for condensation. These results suggest the potential use of JAZ2 condensates to construct artificial membraneless organelles in plant cells.
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The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We thank Drs. M. Matsuoka and H. Yoshioka for their helpful suggestions in transient assays using rice protoplasts and N. benthamiana, respectively, H. Shibata and T. Kojima for useful suggestions on FRAP and SLiCE techniques, respectively, T. Nakagawa, C. Dean, and Y. Habu for the gifts of their plasmids used for the preparation of the Gateway-based vectors pUGW42 and pUGW45, the pETL8 based plasmids, carrying MBP–GFP, MBP–GFP–FCA–RRM and MBP–GFP–FCA–PrLD, and pGFP–Ex–ENS, respectively, C. Ueguchi in Nagoya University for helpful discussion, and Editage (www.editage.com) for English language editing.
Funding
This work was partially supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI (JP16K08140 and JP22K05427) and Nagoya University–National Institute of Advanced Industrial Science and Technology (NU–AIST) alliance project. YK was supported by the Japan Science and Technology Agency (JST) SPRING (JPMJSP2125) and “Graduate Program of Transformative Chem-Bio Research” in Nagoya University, supported by The Ministry of Education, Culture, Sports, Science and Technology (MEXT) (WISE Program).
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All authors contributed to the study conception and/or design. Material preparation, data collection, and analysis were performed by YK, YJ, TA, YY, HG, NH, and HK. OsJAZ2 condensation in Oryza sativa cells was first reported by HK. Data interpretation was performed by TH and ST in addition to the authors described above. The first draft of the manuscript was written by YK and ST and improved by the other authors’ suggestions. All authors read and approved the final manuscript.
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Koja, Y., Joshima, Y., Yoritaka, Y. et al. Formation of subcellular compartments by condensation-prone protein OsJAZ2 in Oryza sativa and Nicotiana benthamiana leaf cells. Plant Cell Rep 42, 269–286 (2023). https://doi.org/10.1007/s00299-022-02955-x
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DOI: https://doi.org/10.1007/s00299-022-02955-x