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Molecular crowding facilitates bundling of IMPDH polymers and cytoophidium formation

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Abstract

The cytoophidium is a unique type of membraneless compartment comprising of filamentous protein polymers. Inosine monophosphate dehydrogenase (IMPDH) catalyzes the rate-limiting step of de novo GTP biosynthesis and plays critical roles in active cell metabolism. However, the molecular regulation of cytoophidium formation is poorly understood. Here we show that human IMPDH2 polymers bundle up to form cytoophidium-like aggregates in vitro when macromolecular crowders are present. The self-association of IMPDH polymers is suggested to rely on electrostatic interactions. In cells, the increase of molecular crowding with hyperosmotic medium induces cytoophidia, while the decrease of that by the inhibition of RNA synthesis perturbs cytoophidium assembly. In addition to IMPDH, CTPS and PRPS cytoophidium could be also induced by hyperosmolality, suggesting a universal phenomenon of cytoophidium-forming proteins. Finally, our results indicate that the cytoophidium can prolong the half-life of IMPDH, which is proposed to be one of conserved functions of this subcellular compartment.

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Acknowledgements

We thank the Molecular and Cell Biology Core Facility (MCBCF) at the School of Life Science and Technology, ShanghaiTech University for providing technical support.

Funding

This work was supported by grants from Ministry of Science and Technology of China (No. 2021YFA0804700), National Natural Science Foundation of China (No. 31771490), Shanghai Science and Technology Commission (No. 20JC1410500) and the UK Medical Research Council (grant nos. MC_UU_12021/3 and MC_U137788471) for grants to J.L.L.

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  1. Chia-Chun Chang

    Present address: Institute of Biotechnology, National Taiwan University, Taipei, 106, Taiwan

Authors and Affiliations

  1. School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, China

    Chia-Chun Chang, Jiale Zhong, Ziheng Zhang, Gerson Dierley Keppeke & Ji-Long Liu

  2. Institute of Biotechnology, National Taiwan University, Taipei, 106, Taiwan

    Min Peng & Li-Ying Sung

  3. Rheumatology Division, Escola Paulista de Medicina, Universidade Federal de Sao Paulo, Sao Paulo, SP, 04023-062, Brazil

    Gerson Dierley Keppeke

  4. Agricultural Biotechnology Research Center, Academia Sinica, Taipei, 115, Taiwan

    Li-Ying Sung

  5. Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, OX1 3PT, UK

    Ji-Long Liu

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Contributions

CCC conceived and designed the studies, analyzed the data. CCC, MP, JZ, ZZ and GDK performed the experiments. JLL and LYS supervised the project. CCC and JLL wrote and edited the manuscript.

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Correspondence to Ji-Long Liu.

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18_2022_4448_MOESM1_ESM.jpg

Supplementary file1 Figure S1. Hyperosmotic medium does not induce the aggregation of PAICS, GMPS and HPRT. Immunofluorescence of wild-type HEK 293T cells treated with 300 mM sucrose for 3 hours. No detectable aggregates of PAICS, GMPS or HPRT were observed in the cells. Scale bars = 20 μm. (JPG 994 kb)

Supplementary file2 Movie S1. Live-cell imaging showing the dynamics of IMPDH amorphous clumps. OFP-IMPDH2 expressing HeLa cells were treated with MPA. Time intervals of each frame is 40 seconds. (AVI 59 kb)

Supplementary file3 Movie S2. Live-cell imaging showing an IMPDH amorphous clump transforms in association with the ER. GFP-Sec61β (green) and OFP-IMPDH2 (red) expressing HeLa cells were treated with MPA. Time intervals of each frame is 5 seconds. (AVI 688 kb)

Supplementary file4 Movie S3. Live-cell imaging showing the transformation of IMPDH amorphous clumps to filaments. OFP-IMPDH2 expressing HeLa cells were treated with MPA. Time intervals of each frame is 60 seconds. (AVI 100 kb)

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Chang, CC., Peng, M., Zhong, J. et al. Molecular crowding facilitates bundling of IMPDH polymers and cytoophidium formation. Cell. Mol. Life Sci. 79, 420 (2022). https://doi.org/10.1007/s00018-022-04448-2

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