The<i>O</i>-GlcNAc transferase OGT is a conserved and essential regulator of the cellular and organismal response to hypertonic stress

Urso, Sarel J.; Comly, Marcella E.; Hanover, John A.; Lamitina, Todd

doi:10.1101/2020.05.01.072033

Cited by 5 publications

(17 citation statements)

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“…1C) that recruit OGT to specific substrates (3,(30)(31)(32)(33)(34). Recent evidence suggests OGT can regulate Caenorhabditis elegans physiology independently of its catalytic functions (35,36); however, given that OGT is not essential in C. elegans (37), it remains unclear whether OGT's binding interactions are essential as well as whether they broadly regulate mammalian physiology independent of OGT's enzymatic activity (38,39).…”

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confidence: 99%

Mammalian cell proliferation requires noncatalytic functions of O-GlcNAc transferase

Levine

Potter

Joiner

et al. 2021

Proc. Natl. Acad. Sci. U.S.A.

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O-GlcNAc transferase (OGT), found in the nucleus and cytoplasm of all mammalian cell types, is essential for cell proliferation. Why OGT is required for cell growth is not known. OGT performs two enzymatic reactions in the same active site. In one, it glycosylates thousands of different proteins, and in the other, it proteolytically cleaves another essential protein involved in gene expression. Deconvoluting OGT’s myriad cellular roles has been challenging because genetic deletion is lethal; complementation methods have not been established. Here, we developed approaches to replace endogenous OGT with separation-of-function variants to investigate the importance of OGT’s enzymatic activities for cell viability. Using genetic complementation, we found that OGT’s glycosyltransferase function is required for cell growth but its protease function is dispensable. We next used complementation to construct a cell line with degron-tagged wild-type OGT. When OGT was degraded to very low levels, cells stopped proliferating but remained viable. Adding back catalytically inactive OGT rescued growth. Therefore, OGT has an essential noncatalytic role that is necessary for cell proliferation. By developing a method to quantify how OGT’s catalytic and noncatalytic activities affect protein abundance, we found that OGT’s noncatalytic functions often affect different proteins from its catalytic functions. Proteins involved in oxidative phosphorylation and the actin cytoskeleton were especially impacted by the noncatalytic functions. We conclude that OGT integrates both catalytic and noncatalytic functions to control cell physiology.

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confidence: 99%

Mammalian cell proliferation requires noncatalytic functions of O-GlcNAc transferase

Levine

Potter

Joiner

et al. 2021

Proc. Natl. Acad. Sci. U.S.A.

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“…In the same genetic screen that identified cpf-2 and symk-1 , we also identified null alleles in the O-GlcNAc transferase ogt-1 (U rso et al . 2020).…”

Section: Discussionmentioning

confidence: 83%

“…However, evidence from thousands of RNAseq datasets does not reveal evidence for APA of gpdh-1 itself (S teber et al . 2019) and genetic studies show that gpdh-1 null alleles can still activate hypertonic adaptation (U rso et al . 2020).…”

Section: Discussionmentioning

confidence: 99%

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

Lamitina

Urso

Sathaseevan

et al. 2023

Preprint

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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 inC. elegansfor mutants with no induction of osmolyte biosynthesis gene expression (Nio mutants). Mutations in twoniogenes encoded missense mutation incpf-2/CstF64 andsymk-1/Symplekin. Bothcpf-2andsymk-1are nuclear components of the highly conserved 3 mRNA cleavage and polyadenylation complex.cpf-2andsymk-1block the hypertonic induction ofgpdh-1and other osmotically induced mRNAs, suggesting they act at the transcriptional level. We generated a functional auxin-inducible degron (AID) allele forsymk-1and found that acute, post-developmental degradation in the intestine and hypodermis was sufficient to cause the Nio phenotype.symk-1andcpf-2exhibit 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-2andsymk-1specifically affect the osmotic stress response since heat shock-induced upregulation of ahsp-16.2::GFPreporter 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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“…In addition to catalyzing both glycosylation and cleavage reactions, OGT interacts with binding partners ( Figure 1C) that recruit OGT to specific substrates (3,(30)(31)(32)(33)(34). Recent evidence suggests OGT may regulate C. elegans physiology independent of its catalytic function (36,37); however, given that OGT is not essential in C.…”

Section: Introductionmentioning

confidence: 99%

Mammalian Cell Proliferation Requires Noncatalytic Functions of O-GlcNAc Transferase

Levine

Potter

Joiner

et al. 2020

Preprint

View full text Add to dashboard Cite

O-GlcNAc transferase (OGT), found in the nucleus and cytoplasm of all mammalian cell types, is essential for cell proliferation. Why OGT is required for cell growth is not known. OGT performs two enzymatic reactions in the same active site. In one, it glycosylates thousands of different proteins, and in the other, it proteolytically cleaves another essential protein involved in gene expression. Deconvoluting OGT's myriad cellular roles has been challenging because genetic deletion is lethal; complementation methods have not been established. Here, we developed approaches to replace endogenous OGT with separation-of-function variants to investigate the importance of OGT's enzymatic activities for cell viability. Using genetic complementation, we found that OGT's glycosyltransferase function is required for cell growth but its protease function is dispensable. We next used complementation to construct a cell line with degron-tagged wild-type OGT. When OGT was degraded to very low levels, cells stopped proliferating but remained viable. Adding back catalytically-inactive OGT rescued growth. Therefore, OGT has an essential noncatalytic role that is necessary for cell proliferation. By developing a method to quantify how OGT's catalytic and noncatalytic activities affect protein abundance, we found that OGT's noncatalytic functions often affect different proteins from its catalytic functions. Proteins involved in oxidative phosphorylation and the actin cytoskeleton were especially impacted by the noncatalytic functions. We conclude that OGT integrates both catalytic and noncatalytic functions to control cell physiology.

show abstract

TheO-GlcNAc transferase OGT is a conserved and essential regulator of the cellular and organismal response to hypertonic stress

Cited by 5 publications

References 49 publications

Mammalian cell proliferation requires noncatalytic functions of O-GlcNAc transferase

Mammalian cell proliferation requires noncatalytic functions of O-GlcNAc transferase

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

Mammalian Cell Proliferation Requires Noncatalytic Functions of O-GlcNAc Transferase

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