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bioRxiv · 10.1101/2023.01.23.525244

Regulation of the hypertonic stress response by the 3' mRNA cleavage and polyadenylation complex

Lamitina, T.·Urso, S.·Sathaseevan, A.·Derry, W. B.
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Abstract

Maintenance of osmotic homeostasis is one of the most aggressively defended homeostatic setpoints in physiology. One major mechanism of osmotic homeostasis involves the upregulation of proteins that catalyze the accumulation of solutes called organic osmolytes. To better understand how osmolyte accumulation proteins are regulated, we conducted forward genetic screen in C. elegans for mutants with no induction of osmolyte biosynthesis gene expression (Nio mutants). nio-3 mutants encoded a missense mutation in cpf-2/CstF64 while nio-7 mutants encoded a missense mutation in symk-1/Symplekin. Both cpf-2 and symk-1 are nuclear components of the highly conserved 3 mRNA cleavage and polyadenylation complex. cpf-2 and symk-1 block the hypertonic induction of gpdh-1 and other osmotically induced mRNAs, suggesting they act at the transcriptional level. We generated a functional auxin-inducible degron (AID) allele for symk-1 and found that acute, post-developmental degradation in the intestine and hypodermis was sufficient to cause the Nio phenotype. symk-1 and cpf-2 exhibit genetic interactions that strongly suggest they function through alterations in 3 mRNA cleavage and/or alternative polyadenylation. Consistent with this hypothesis, we find that inhibition of several other components of the mRNA cleavage complex also cause a Nio phenotype. cpf-2 and symk-1 specifically affect the osmotic stress response since heat shock-induced upregulation of a hsp-16.2::GFP reporter is normal in these mutants. Our data suggest a model in which alternative polyadenylation of one or more mRNAs is essential to regulate the hypertonic stress response.

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Lamitina, T., Urso, S., Sathaseevan, A., Derry, W. B.. 2023-01-24. Regulation of the hypertonic stress response by the 3' mRNA cleavage and polyadenylation complex. https://doi.org/10.1101/2023.01.23.525244

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