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PUS7 cytoplasmic localization directs a pseudouridine-mediated cellular stress response

ABSTRACT: Pseudouridine (Ψ) is an abundant post-transcriptional modification found across all classes of RNA. Since its discovery in mRNAs a decade ago, it has been widely speculated that Y might govern additional post-transcriptional regulation of gene expression. Here, we demonstrate that one of the principal enzymes responsible for adding Y to mRNAs, pseudouridine synthase 7 (PUS7), accumulates in the cytoplasm under a variety of stress conditions in Saccharomyces cerevisiae and BEAS-2B human epithelial lung cells. The localization of PUS7 to the cytoplasm promotes Y-incorporation into hundreds of different mRNA sequences and increases cellular fitness under ROS and divalent metal ion stress. The preponderance of newly identified Y-sites lies within portions of the mRNA important for post-transcriptional control—-coding regions and 3’ UTRs. Quantitative proteomics reveal that shifts in the cellular post-transcriptional modification landscape upon PUS7 relocalization reshapes the proteome. Our data suggest a mechanism whereby stressors localize PUS7 in the cytoplasm to enable the direct modification and regulation of stress response mRNAs, thereby protecting cells from further stress-induced damage. (2A-PUS7-control682832B-PUS7-control682842C-PUS7-control682854A-PUS7-NES-Co682864A-PUS7-NES-Co682874B-PUS7-NES-Co682885A-PUS7-control-Co682895B-PUS7-control-Co682905C-PUS7-control-Co68291PUS7-NES_CU_1A 75059PUS7-NES_CU_1B 75060PUS7-NES_CU_1C 75061PUS7-NLS_CU_2A 75062PUS7-NLS_CU_2B 75063PUS7-NLS_CU_2C 75064PUS7-WT_CU_3A 75065PUS7-WT_CU_3B 75066PUS7-WT_CU_3C 75067)

INSTRUMENT(S):

ORGANISM(S): Saccharomyces Cerevisiae By4741

SUBMITTER: Minli Ruan

LAB HEAD: Kristin Koutmou

PROVIDER: PXD052684 | Pride | 2026-01-28

REPOSITORIES: Pride

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			Action	DRS
	MSB11373.mztab	Mztab
	MSB11373APool_30MinDIA113023GPF_01.mzML	Mzml
	MSB11373APool_30MinDIA113023GPF_02.mzML	Mzml
	MSB11373APool_30MinDIA113023GPF_03.mzML	Mzml
	MSB11373APool_30MinDIA113023GPF_04.mzML	Mzml

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Project description:In vitro maturation (IVM) oocyte is a key component of assisted reproduction in pigs. Asynchronous nucleoplasmic maturation and incomplete cytoplasmic maturation is one of the key factors for impaired maturation and quality of oocytes matured in vitro. Although Single-omics techniques have been applied to the study of porcine oocyte maturation mechanisms in the past, it is difficult to address a systematic and comprehensive explanation of the complex maturation regulatory network through Single-omics data alone, and it is insufficient to account for the chain of transmission of the regulation of genetic information expression. This is the first study which determine the gene transcripts, host potential proteins interactions, and key functional metabolites involved in the maturation mechanism of porcine oocytes. We found that among 2624 and 124 differentially expressed genes and proteins respectively, genes DNMT1, HENMT1, H1FOO, GDF9, NUP214, CAST and GSTM3 were differentially coexisted at both levels，these differentially expressed genes are closely associated with 13 differentially expressed metabolites.In addition, the genes DNMT1 , HENMT1, H1FOO, GDF9, NUP214, CAST and GSTM3 were differentially expressed at both transcript and protein levels. To address further functional analysis, we conducted GO and KEGG analysis at mRNAs and metabolites level to outcome the involvement of multiple epigenetic regulatory mechanisms affecting nuclear and cytoplasmic maturation of porcine oocytes. Thus, we systematically illustrated how epigenetic modifications affect nuclear and cytoplasmic maturation of porcine oocytes from the dimension of the transmission chain of genetic information expression regulation. Specifically, it includes: 1) .The increase in DNA methylation level and the decrease in histone acetylation level lead to the reduction of chromatin accessibility, thereby triggering global transcriptional silencing. 2).A large number of mRNAs that have been transcribed in the nucleus need to undergo complex modifications before they can be recognized by the nuclear pore complex (NPC) and transported to the cytoplasm. 3). The mRNAs transported to the cytoplasm are selectively translated activated according to the requirements of the cell maturation process. This spatiotemporal heterogeneity of translation activation is strictly regulated by post-transcriptional modifications. 4 ). Post-translational modifications regulate the active sialic acid metabolism to prepare for the fertilization process; the macro and micro changes in glutathione metabolism and mitochondrial metabolism aim to meet the dynamic energy requirements of the cell and regulate intracellular redox homeostasis to reduce oxidative damage.

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PUS7 cytoplasmic localization directs a pseudouridine-mediated cellular stress response

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