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PUS10-induced tRNA fragmentation impacts retrotransposon-driven inflammation

ABSTRACT: Pseudouridine synthases (PUSs) catalyze the isomerization of uridine (U)-to-pseudouridine (Ψ) and have emerging roles in development and disease. How PUSs adapt gene expression under stress remains mostly unexplored. We identify an unconventional role for the Ψ “writer” PUS10 impacting intracellular innate immunity. Using Pus10 knock-out mice, we uncover cell-intrinsic upregulation of interferon (IFN) signalling, conferring resistance to inflammation in vivo. Pus10 loss alters tRNA-derived small RNAs (tdRs) abundance, perturbing translation and endogenous retroelements expression. We found one of the differentially expressed tDRs is derived from tRNA glycine (tDR Gly). To delinate the tDR Gly interactome, we employed an optimized pull-down strategy using PUS10 KO cells followed by data-independent acquisition-based quantitative mass spectrometry analysis.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture, Fibroblast

SUBMITTER: Sudip Ghosh

LAB HEAD: Jenny Hansson

PROVIDER: PXD061235 | Pride | 2025-05-27

REPOSITORIES: Pride

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			Action	DRS
	20241020_125149_MEFs_CoIP_PUS10KO_directDIA_SN18_241020.sne	Other
	241018_MEFs_CoIP_PUS10KO_glybio_1_120min_Exploris_FAIMS_DIA_CV-45-60.raw	Raw
	241018_MEFs_CoIP_PUS10KO_glybio_2_120min_Exploris_FAIMS_DIA_CV-45-60.raw	Raw
	241018_MEFs_CoIP_PUS10KO_glybio_3_120min_Exploris_FAIMS_DIA_CV-45-60.raw	Raw
	241018_MEFs_CoIP_PUS10KO_glybio_4_120min_Exploris_FAIMS_DIA_CV-45-60.raw	Raw

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Project description:Parental dietary conditions can influence the metabolic traits of offspring. In mice, paternal consumption of low protein diet alters cholesterol and lipid metabolism of progeny. Here, we examine RNA species expressed in male reproductive tissues of mice. Protein restriction leads to altered levels of multiple small RNAs in mature sperm, as well as throughout the male reproductive tract, with decreased levels of let-7 family members and increased levels of 5â?? fragments of tRNA-Gly isoacceptors. Intriguingly, tRNA fragments are scarce in the testis, but their levels increase in sperm during post-testicular maturation in the epididymis. We find that epididymosomes â?? extracellular vesicles which fuse with sperm during epididymal transit â?? exhibit RNA payloads closely matching those of mature sperm, and can deliver tRNA fragments to immature sperm in vitro both in mouse and in bull. Finally, we show that tRNA-Gly-GCC fragments play a role in repressing genes associated with the endogenous retroelement MERVL, both in ES cells and in preimplantation embryos. Our results shed light on small RNA biogenesis during post-testicular sperm maturation, and link tRNA fragments to regulation of endogenous retroelements active in the early embryo. Zygotes were generated by IVF from animals fed a control diet. These embryos were then microinjected with various combinations of small RNAs and control RNA (HIS3.3::GFP). Follwoing injections the zygotes were developed and allowed to develop until 2 cell (2C) or 4 cell (4C) stage when single embryo RNA seq was carried out to study gene expression changes

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