The CCT chaperonin and actin modulate the ER and RNA-binding protein condensation during oogenesis to maintain translational repression of maternal mRNA and oocyte quality

Elaswad, M. T.; Gao, M.; Tice, V. E.; Bright, C. G.; Thomas, G. M.; Munderloh, C.; Trombley, N. J.; Haddad, C. N.; Johnson, U. G.; Cichon, A. N.; Schisa, J. A.

doi:10.1101/2024.07.01.601596

bioRxiv · 10.1101/2024.07.01.601596

The CCT chaperonin and actin modulate the ER and RNA-binding protein condensation during oogenesis to maintain translational repression of maternal mRNA and oocyte quality

Elaswad, M. T.·Gao, M.·Tice, V. E.·Bright, C. G.·Thomas, G. M.·Munderloh, C.·Trombley, N. J.·Haddad, C. N.·Johnson, U. G.·Cichon, A. N.·Schisa, J. A.

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Abstract

The regulation of maternal mRNAs is essential for proper oogenesis, the production of viable gametes, and to avoid birth defects and infertility. Many oogenic RNA-binding proteins have been identified with roles in mRNA metabolism, some of which localize to dynamic ribonucleoprotein granules and others that appear dispersed. Here, we use a combination of in vitro condensation assays and the in vivo C. elegans oogenesis model to determine the intrinsic properties of the conserved KH-domain MEX-3 protein and to identify novel regulators of MEX-3 and the Lsm protein, CAR-1. We demonstrate that MEX-3 undergoes liquid-liquid phase separation and appears to have intrinsic gel-like properties in vitro. We also identify novel roles for the CCT chaperonin and actin in preventing ectopic RNA-binding protein condensates in maturing oocytes that appear to be independent of MEX-3 folding. CCT and actin also oppose the expansion of ER sheets that may promote ectopic condensation of RNA-binding proteins that are associated with de-repression of maternal mRNA. This regulatory network is essential to preserve oocyte quality, prevent infertility, and may have implications for understanding the role of hMex3 phase transitions in cancer. Significance statementO_LIThe molecular mechanisms that regulate phase transitions of oogenic RNA-binding proteins are critical to elucidate but are not fully understood. C_LIO_LIWe identify novel regulators of RNA-binding protein phase transitions in maturing oocytes that are required to maintain translational repression of maternal mRNAs and oocyte quality. C_LIO_LIThis study is the first to elucidate a regulatory network involving the CCT chaperonin, actin, and the ER for phase transitions of RNA-binding proteins during oogenesis. Our findings for the conserved MEX-3 protein may also be applicable to better understanding the role of hMex3 phase transitions in cancer. C_LI

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Elaswad, M. T., Gao, M., Tice, V. E., Bright, C. G., Thomas, G. M., Munderloh, C., Trombley, N. J., Haddad, C. N., Johnson, U. G., Cichon, A. N., Schisa, J. A.. 2024-07-04. The CCT chaperonin and actin modulate the ER and RNA-binding protein condensation during oogenesis to maintain translational repression of maternal mRNA and oocyte quality. https://doi.org/10.1101/2024.07.01.601596

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