Dataset Information

Dissecting the Molecular Crosstalk between O-GlcNAcylation and Phosphorylation via Quantitative Proteomics and Phosphoproteomics

ABSTRACT: In this study, we explored the cross-talk between phosphorylation and O-GlcNAcylation using the pancreatic ductal cell line PANC-1 as a model. Proteome and phosphoproteome changes were measured for cells treated with OSMI-1, a specific inhibitor to O-GlcNAc transferase (OGT) and Thiamet G, a specific inhibitor to O-GlcNAcase (OGA). Among the 8938 phosphorylation sites quantified, 1399 phosphosites on 870 proteins and 1480 phosphosites on 896 proteins were significantly altered by OSMI-1 and TMG treatment, respectively, demonstrating extensive cross-talk between O-GlcNAcylation and phosphorylation. In-depth analysis revealed widespread phosphorylation changes of the kinome and phosphatome, even after a short-term perturbation with inhibitors to O-GlcNAc cycling enzymes. Notably, we identified that phosphorylation of OGA at S364 is specifically mediated by casein kinase 2 alpha (CK2α), which was supported by phosphoproteomic profiling, kinase inhibition experiments, and in vitro kinase assays. These results uncover a regulatory mechanism underlying glycosylation-dependent cellular signaling through the interplay of phosphorylation and O-GlcNAcylation.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

DISEASE(S): Carcinoma

SUBMITTER: Ci Wu

LAB HEAD: Junfeng Ma

PROVIDER: PXD070284 | Pride | 2026-06-29

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20240523_111741_pST_TMG_OSMI_DMSO_Report_PTM.csv	Csv
	20240523_111741_pST_TMG_OSMI_DMSO_Report_normal.csv	Csv
	20240523_111741_pST_TMG_OSMI_DMSO_Report_peptide.csv	Csv
	20240523_111741_pST_TMG_OSMI_DMSO_Report_protein.csv	Csv
	Panc1_pST_TiO2_DMSO_DIA1new.raw	Raw

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Publications

Deciphering O‑GlcNAc-Dependent Signaling Via Integrated Proteomics and Phosphoproteomics.

Wu Ci C Hou Chunyan C Wang Xinyue X Peng Yihan Y Lin Yao Y Byers Stephen W SW Pei Huadong H Ma Junfeng J

ACS omega 20260604 23

Post-translational modifications (PTMs) on proteins play crucial roles in various biological processes. Two highly dynamic modifications, phosphorylation and O-linked N-acetylglucosamine modification (O-GlcNAcylation), are essential for cellular physiology and pathology. Emerging evidence suggests intimate crosstalk between phosphorylation and O-GlcNAcylation on multiple proteins. However, the precise nature of their crosstalk remains largely unknown. In this study, we explored the crosstalk bet ...[more]

PMID: 42326659

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Project description:Drosophila development is a complex and dynamic process regulated, in part, by members of the Polycomb (Pc), Trithorax (Trx) and Compass chromatin modifier complexes. O-GlcNAc Transferase (OGT/SXC) is essential for Pc repression suggesting that the O-GlcNAcylation of proteins plays a key role in regulating development. OGT transfers N-acetyl-D-glucosamine (GlcNAc) onto hydroxyl groups of serine or threonine residues of key transcriptional regulators using the nutrient-derived UDP-GlcNAc as a substrate, which is dynamically removed by O-GlcNAcase (OGA). We performed ChIP-chip and microarray analysis after OGT or OGA RNAi knockdown in Drosophila S2 cells and found that O-GlcNAc was elevated genome wide particularly at genes related to mitosis and cell cycle in OGA RNAi cells, but not at sites co-occupied by Pc member Pleiohomeotic (Pho), such as the Hox and NK homeobox gene clusters. Microarray analysis suggested that altered O-GlcNAc cycling perturbed the expression of genes associated with morphogenesis and cell cycle regulation. To examine the in vivo consequences of disturbed O-GlcNAc cycling in the whole animal, we produced a null allele of oga (ogadel.1) in Drosophila. Epigenetic activators including Trx group members Trithorax (Trx), Absent small or homeotic discs 1 (Ash1) and Compass member Set1 histone methyltransferases are O-GlcNAc modified in ogadel.1 mutants. ogadel.1 mutants displayed altered expression of a distinct set of cell cycle related genes in ovaries. Our results suggest that the loss of OGA could affect epigenetic machinery by accumulating O-GlcNAc on numerous chromatin factors including Trx, Ash1 and Set1 in Drosophila. We performed affymetrix tilingarray analysis after OGT or OGA RNAi knockdown in Drosophila S2 cells to find if that O-GlcNAc was elevated genome wide particularly at genes related to mitosis and cell cycle in OGA RNAi cells, but not at sites co-occupied by Pc member Pleiohomeotic (Pho), ------------------------------- This represents the gene expression component only