Pseudouridine guides germline small RNA transport and epigenetic inheritance

Herridge, R. P.; Dolata, J.; Migliori, V.; de Santis Alves, C.; Borges, F.; Van Ex, F.; Lin, A.; Bajczyk, M.; Leonardi, T.; Hendrick, A.; Schorn, A. J.; Kouzarides, T.; Martienssen, R. A.

doi:10.1101/2023.05.27.542553

bioRxiv · 10.1101/2023.05.27.542553

Pseudouridine guides germline small RNA transport and epigenetic inheritance

Herridge, R. P.·Dolata, J.·Migliori, V.·de Santis Alves, C.·Borges, F.·Van Ex, F.·Lin, A.·Bajczyk, M.·Leonardi, T.·Hendrick, A.·Schorn, A. J.·Kouzarides, T.·Martienssen, R. A.

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Abstract

Epigenetic modifications that arise during plant and animal development, such as DNA and histone modification, are mostly reset during gamete formation, but some are inherited from the germline including those marking imprinted genes1. Small RNAs guide these epigenetic modifications, and some are also inherited by the next generation2,3. In C. elegans, these inherited small RNAs have poly (UG) tails4, but how inherited small RNAs are distinguished in other animals and plants is unknown. Pseudouridine ({Psi}) is the most abundant RNA modification but has not been explored in small RNAs. Here, we develop novel assays to detect {Psi} in short RNA sequences, demonstrating its presence in mouse and Arabidopsis microRNAs and their precursors. We also detect substantial enrichment in germline small RNAs, namely epigenetically activated siRNAs (easiRNAs) in Arabidopsis pollen, and piwi-interacting piRNAs in mouse testis. In pollen, pseudouridylated easiRNAs are localized to sperm cells, and we found that PAUSED/HEN5 (PSD), the plant homolog of Exportin-t, interacts genetically with {Psi} and is required for transport of easiRNAs into sperm cells from the vegetative nucleus. We further show that Exportin-t is required for the triploid block: chromosome dosage-dependent seed lethality that is epigenetically inherited from pollen. Thus, {Psi} has a conserved role in marking inherited small RNAs in the germline. One-Sentence SummaryPseudouridine marks germline small RNAs in plants and mammals, impacting epigenetic inheritance via nuclear transport.

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Herridge, R. P., Dolata, J., Migliori, V., de Santis Alves, C., Borges, F., Van Ex, F., Lin, A., Bajczyk, M., Leonardi, T., Hendrick, A., Schorn, A. J., Kouzarides, T., Martienssen, R. A.. 2023-05-30. Pseudouridine guides germline small RNA transport and epigenetic inheritance. https://doi.org/10.1101/2023.05.27.542553

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