Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Spatiotemporal control of subcellular O-GlcNAc signaling using light-inducible Opto-OGT

ABSTRACT: The posttranslational modification of intracellular proteins by O-linked β-N-acetylglucosamine (O-GlcNAc) is a universal regulatory mechanism in multicellular organisms driven by O-GlcNAc transferase (OGT). The complex network of signaling proteins that are activated and involved in nutrient and stress sensing pathways have made it challenging to disentangle the direct role of OGT and O-GlcNAcylation. Here, we describe Opto-OGT, an optogenetic approach that allows precise activation of OGT with light. This tool leverages on photosensitive cryptochrome fused to OGT and potentially blocks the OGT substrate site in dark state. We elucidate how activation of OGT alone can specify for distinct physiological conditions:- a short 15-min OGT activation pulse results in mTORC pathway activation and AMPK suppression, mimicking cellular signaling events activated by glucose sensing. Opto-OGT activation displays a strong temporal dependence as well - switching on OGT for 1 hour results in vice versa that represents nutrient deprivation-like response. Moreover, Opto-OGT can be customized to be activated at specific subcellular locations. As a proof-of-concept, we demonstrate downregulation of site-specific AKT phosphorylation and signaling outputs as a result of OGT recruitment towards the plasma membrane in the presence of insulin. The ability to isolate OGT as a signaling node and uncover its inherent signaling properties enables the dissection of the role of O-GlcNAcylation in whole-cell or specific compartments.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Weiping Han and Xiaoyong Yang

PROVIDER: PXD048816 | JPOST Repository | Thu Jan 23 00:00:00 GMT 2025

REPOSITORIES: jPOST

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	20190611_Human_UniProt.fasta	Fasta
	20220819_JW_QX_14.raw	Raw
	20220819_JW_QX_18.raw	Raw
	20220819_JW_QX_21.raw	Raw
	20220819_JW_QX_24.raw	Raw

Items per page:

1 - 5 of 10

Similar Datasets

Regulation of NRF2 signaling by O-GlcNAcylation of KEAP1

Project description:O-GlcNAcylation is an essential, nutrient-sensitive post-translational modification, but its biochemical and phenotypic effects remain incompletely understood. To address this knowledge gap, we investigated the global transcriptional response to perturbations in O-GlcNAcylation. Unexpectedly, many transcriptional effects of O-GlcNAc transferase (OGT) inhibition were due to the activation of NRF2, the master regulator of redox stress tolerance. Moreover, we found that a signature of low OGT activity strongly correlates with NRF2 activation in multiple tumor expression datasets. Guided by this information, we identified KEAP1 (also known as KLHL19), the primary negative regulator of NRF2, as a direct substrate of OGT. We show that O-GlcNAcylation of KEAP1 at serine 104 is required for the efficient ubiquitination and degradation of NRF2. Interestingly, O-GlcNAc levels and NRF2 activation co-vary in response to glucose fluctuations, indicating that KEAP1 O-GlcNAcylation links nutrient sensing to downstream stress resistance. Our results reveal a novel regulatory connection between nutrient-sensitive glycosylation and NRF2 signaling, and provide a blueprint for future approaches to discover functionally important O-GlcNAcylation events on other KLHL family proteins in various experimental and disease contexts.

2017-06-26 | GSE81740 | GEO

O-GlcNAcylation of Raptor transduces glucose signals to mTORC1

Project description:Complex molecular mechanisms ensure cellular homeostasis between nutrient sensing and energy metabolism. Strong evidence from many laboratories supports a role for the hexosamine biosynthetic pathway (HBP) and its end product, UDP-GlcNAc, as important nutrient sensors. Isotopic photocleavable tagging for O-GlcNAc profiling (isoPTOP) was performed for quantitative and site specific identification of O-GlcNAcylation upon glucose mediated nutrient fluctuation. Mammalian target of Rapamycin complex 1（mTORC1）is a metabolic hub, which can sense the availability of various nutrients. We found that Raptor, the scaffold of the mTORC1 complex, is O-GlcNAcylated at T700. Our finding provides a novel mechanism that UDP-GlcNAc mediates glucose signal to mTORC1 by O-GlcNAcylating Raptor at T700 to regulate mTORC1 activity and cell growth.

2023-08-03 | PXD039536 | Pride

Protein O-GlcNAcylation coupled to Hippo signaling drives vascular dysfunction in diabetic retinopathy [9A]

Project description:Metabolic disorder is emerging as a crucial contributor to the pathogenesis of diabetic vascular complications including diabetic retinopathy (DR), the leading cause of blindness in the working-age population. Yet, the underlying molecular mechanisms of how the disturbed metabolic homeostasis contribute to vascular dysfunction in DR remain elusive. O-GlcNAcylation modification act as a nutrient sensor particularly sensitive to ambient glucose. Here, we observed pronounced O-GlcNAc elevation in retina endothelial cells (ECs) of DR patients and mouse models. Endothelial-specific depletion or pharmacological inhibition of O-GlcNAc transferase efficiently mitigated vascular dysfunctions. Mechanistically, we found that YAP/TAZ, key effectors of the Hippo pathway, are O-GlcNAcylated in DR. We identified Thr383 as an O-GlcNAc site on YAP, which inhibits its phosphorylation at Ser397, leading to its stabilization and activation. Consequently, highly activated YAP/TAZ, promotes vascular dysfunction by inducing a pro-angiogenic and glucose metabolic transcriptional program. These findings emphasize the critical role of O-GlcNAc-Hippo axis in DR pathogenesis and suggest its unique potential as a therapeutic target.

2024-09-24 | GSE266072 | GEO

Protein O-GlcNAcylation coupled to Hippo signaling drives vascular dysfunction in diabetic retinopathy [3A]

2024-09-24 | GSE266071 | GEO

Title: Site specific O-GlcNAcylation of progesterone receptor (PR) supports PR attenuation of interferon stimulated genes (ISGs) and tumor growth in breast cancer

Project description:Hormone receptor (HR) positive breast cancer, defined by expression of estrogen (ER) and/or progesterone (PR) receptor expression, is the most commonly diagnosed type of breast cancer. PR alters the transcriptional landscape to support tumor growth in concert with or independent of ER. Understanding the mechanisms regulating PR function are critical to developing new strategies to treat HR+ breast cancer. O-GlcNAc is a post-translational modification responsible for nutrient sensing that fine tunes protein function. We have previously reported O-GlcNAcylation on PR. Although PR is heavily post translationally modified, primarily through phosphorylation, specific sites of O-GlcNAcylation on PR, and how they regulate PR action, have not been investigated. Using established PR-expressing breast cancer cell lines, we mapped the sites of O-GlcNAcylation on PR. RNA-sequencing revealed site-specific O-GlcNAcylation of PR is critical for ligand-independent suppression of interferon signaling, a regulatory function of PR previously studied in our lab. Furthermore, O-GlcNAcylation of PR enhances PR-driven tumor growth in vivo. We have delineated one mechanism regulating PR function in breast cancer that impacts tumor growth, and provided additional insight into the mechanism through which PR attenuates interferon signaling.

2024-06-17 | MSV000095057 | MassIVE

Expression from C. elegans L1 animals

Project description:Nutrient-driven O-GlcNAcylation of key components of the transcription machinery may epigenetically modulate gene expression in metazoans. Knockouts of the O-GlcNAc cycling enzymes in C. elegans are viable and fertile, allowing a global analysis of the impact of GlcNAcylation. Whole genome transcriptional profiling of the O-GlcNAc cycling mutants confirmed dramatic deregulation of genes in these key pathways. As predicted, the O-GlcNAc cycling mutants show phenotypically altered lifespan and susceptibility to UV stress. L1 larvae were synchronized by starvation for two days after hatching in sterile buffer. Wild type and mutant animals were collected and total RNA isolated for gene expression analysis

2009-12-31 | E-GEOD-18130 | biostudies-arrayexpress

Expression from C. elegans L4 animals

Project description:Nutrient-driven O-GlcNAcylation of key components of the transcription machinery may epigenetically modulate gene expression in metazoans. Knockouts of the O-GlcNAc cycling enzymes in C. elegans are viable and fertile, allowing a global analysis of the impact of GlcNAcylation. Whole genome transcriptional profiling of the O-GlcNAc cycling mutants confirmed dramatic deregulation of genes in these key pathways. As predicted, the O-GlcNAc cycling mutants show phenotypically altered lifespan and susceptibility to UV stress. Similarly aged L4 animals (20) staged by degree of vulval development were hand picked. Wild type and mutant animals were collected and total RNA isolated for gene expression analysis

2010-01-01 | E-GEOD-18131 | biostudies-arrayexpress

Nutrient Responsive O-GlcNAcylation Dynamically Modulates Galectin 3 Secretion

Project description:Endomembrane glycosylation and cytoplasmic O-GlcNAcylation each play essential roles in nutrient sensing, and in fact, characteristic changes in glycan patterns have been described in disease states such as diabetes and cancer. These changes in glycosylation have important functional roles and can drive disease progression. However, little is known about the molecular mechanisms underlying how these signals are integrated and transduced into biological effects. Galectins are proteins that bind glycans and are secreted by a poorly characterized non-classical secretory mechanism. Once outside the cell, galectins bind to terminal galactose residues of cell surface glycans and modulate numerous extracellular functions like clathrin independent endocytosis (CIE). Originating in the cytoplasm, galectins are predicted substrates for O-GlcNAc addition and removal; and, as we have shown, galectin 3 is a substrate for OGT. This study also shows that galectin 3 secretion is sensitive to changes in O-GlcNAc levels. We determined that there is a significant difference in O-GlcNAcylation status between cytoplasmic and secreted galectin 3. We observed dramatic alterations in galectin 3 secretion in response to nutrient conditions and these changes were dependent on dynamic O-GlcNAcylation. Importantly, we showed that these O-GlcNAc driven alterations in galectin 3 secretion drove changes in CIE. These results indicate that dynamic O-GlcNAcylation of galectin 3 plays a role in modulating its secretion and can tune its function of transducing nutrient sensing information coded in cell surface glycosylation into biological effects.

2022-04-01 | PXD028139 | Pride

Expression from C. elegans L4 animals

2010-01-01 | GSE18131 | GEO

Expression from C. elegans L1 animals

2010-01-01 | GSE18130 | GEO