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Zhang N, Meng Y, Mao S, Ni H, Huang C, Shen L, Fu K, Lv L, Yu C, Meekrathok P, Kuang C, Chen F, Zhang Y, Yuan K. FBXO31-mediated ubiquitination of OGT maintains O-GlcNAcylation homeostasis to restrain endometrial malignancy. Nature communications 2025 16(1) 39894887
Abstract:
Protein O-GlcNAcylation is a post-translational modification coupled to cellular metabolic plasticity. Aberrant O-GlcNAcylation has been observed in many cancers including endometrial cancer (EC), a common malignancy in women. However, clinical characterization of dysregulated O-GlcNAcylation homeostasis in EC and interrogating its molecular mechanism remain incomplete. Here we report that O-GlcNAcylation level is positively correlated with EC histologic grade in a Chinese cohort containing 219 tumors, validated in The Cancer Genome Atlas dataset. Increasing O-GlcNAcylation in patient-derived endometrial epithelial organoids promotes proliferation and stem-like cell properties, whereas decreasing O-GlcNAcylation limits the growth of endometrial cancer organoids. CRISPR screen and biochemical characterization reveal that tumor suppressor F-box only protein 31 (FBXO31) regulates O-GlcNAcylation homeostasis in EC by ubiquitinating the O-GlcNAc transferase OGT. Downregulation of O-GlcNAcylation impedes EC tumor formation in mouse models. Collectively, our study highlights O-GlcNAcylation as a useful stratification marker and a therapeutic vulnerability for the advanced, poorly differentiated EC cases.
O-GlcNAc proteins:
FR1L5, ERIC2, PKHG3, CCD78, ZN320, WDR27, MBPHL, CFA69, CDHR4, PINLY, VWA3A, NPB15, AKD1B, CC154, AGAP5, MET15, SAC31, CB081, TTL13, ENO4, CFA96, ZSWM8, OCAT2, NPIB5, LYRM9, ZBBX, CMI2B, NOTO, ARG33, CMA1B, DCD2C, IQCF6, ZN487, SHSA9, ADM5, PROB1, PTG3L, TM262, CK098, CS084, DCDC1, BACH, S28A1, SYT5, VMA5A, PITM1, MPP10, CCL22, EST2, P3C2B, UBE2C, TOR1A, P5I11, RPC1, PSA7, TR13B, RGS12, HAT1, CE290, NPFF, PGRC2, NPHP1, SIVA, TRI18, HMGB3, MRP3, INSI1, SRGP3, ADCY6, FGF16, IF4G3, ADA20, HTSF1, RAD21, NPA1P, L37A3, EREP2, NMD3B, TSN1, DPM1, TOM1, NSMA, HUS1, CTNS, AOC2, NPHP4, SNN, PROP1, PEX14, MED24, Z324A, UBP2, ZMY10, SPI2, IDHC, RBBP9, SOAT2, SPF27, DNJC8, FGF18, UBP1, BAIP3, FARP2, ZN432, KDM4B, HEXI1, UPP2, REC8, NDUB4, MRP6, RASL1, TACC2, RPGF3, BAG4, ATS2, ACSL3, RAB36, FRITZ, DRC4, CCNE2, ANF, PROC, PDGFA, LCAT, B3A2, CFAI, CO5A2, PGK2, RU2B, FGF4, PKHD1, SPRC, INHBB, F163B, OST4, PRR35, PAL4G, CGB3, VSIG8, COX8A, MYBB, RALA, CD79A, PCNA, ECP, PPA5, PRG2, OXDA, ZNF44, RFA2, UD2B7, STMN1, YBOX3, ZNF19, ZNF23, INAR1, SAP3, PRS6A, GBRB1, TNR1A, AOC1, AMY2B, PSB1, LMNB1, CAN3, CP3A5, HNF1A, CP2A7, ENDOU, ZNF10, DRD4, CCNO, CP7A1, CRYGS, EST1, RIR1, PAR1, MCM3, PSA2, PSA3, HMGB2, KAD4, PSB6, MZF1, RBL1, PTPRM, 3MG, PBLD, STX2, ERD22, MCM5, GALNS, SDC2, BMP8B, SPB6, RFA3, DEK, PRS7, GGT2, TNR3, TALDO, IF4A3, FEN1, HNF4A, M3K8, LAP2A, LAP2B, STAT6, MTHR, AL3B1, S15A1, MSS4, RFX2, INAR2, PRC2A, HSP13, GCST, PAXI, MAPK2, RPIA, CENPA, CENPF, PRI1, ACADV, NUMB, 5NTC, GUAA, DNLI4, ZNF83, IRAK1, CCR8, CLCN7, VAV2, ZN140, BIEA, MK10, OAZ1, AT1B3, DRG2, RAD, FGF8, UCN1, FOXA1, ALR, CTBP2, BCAR1, WDR4, CORO7, FBXW4, CABP4, RAB15, DJB13, SC61G, PTEN, SRP54, CKS1, PFD3, SUMO2, PP1A, PRS4, RUXG, LSM3, TYB4, TRA2B, 3BP2, RBM6, RPP38, HEPC, GPSM2, BAT1, RT35, CBX1, METK1, TF2B, CDK3, E2F1, MLRA, MYL5, ID2, P, HGFA, LGUL, RAD51, MCL1, PDE4C, SG11B, F27E3, DMPK, R51A2, INCA1, CF210, MGAT2, SFSWA, BIRC1, PAFA, IFT88, KLF10, SEM3B, PHLP, PPIG, PEX6, RB33A, DPOA2, SRC8, FLOT2, GALE, SLBP, HABP2, MCM6, TTLL4, RRP5, 2A5D, CND2, SPCS2, AR6P1, K0040, PEA15, CDK10, PON2, 2A5A, IRF4, TAF5, TAF1C, TRBP2, TRIP6, ZNF79, E2F4, ZN239, SNAT, NFE2, STX1A, MEA1, MK06, CDKN3, THTR, PCKGM, ZN827, MOONR, CL16A, MTHSD, FA83E, CF184, TECT1, PRSR3, TXIP1, CCD57, HKDC1, FHOD3, BTBDG, GFOD2, TKFC, TTLL9, DRC12, CDNF, HYDIN, SRCRM, PCDP1, CCD40, IQCC, PLCH1, VAS1L, PDCD4, LRC23, ZC21C, T4S20, FBLN7, CC157, M3K19, PAN3, TRI61, HERC4, MDH1B, ZN311, CC038, DYLT4, MEIG1, RRP12, WDR38, EFHC1, COL12, PA2GC, MANEA, PKHS1, SNPC4, F201A, F120S, MORN1, CFA70, UBX11, AXDN1, MRP7, ZFY27, AGRF1, CFA58, KIF24, SYT6, IER5L, ARM12, KAD9, DAAF9, CF163, RHG40, TMCO4, STPG1, EFCB6, CROCC, MEX3C, CBPC2, MCAF2, GP157, AXA81, CCD30, AGAP6, SLX4I, NXNL2, ARMC3, TTC23, FBX31, FBXW9, AL2CL, KFA, ATG9B, FTM, SLN13, FRPD2, Z518A, DMKN, TTLL5, ABLM2, ELP2, IQCE, CCSAP, ACD10, LCN12, OGFD2, NANP8, SAPL1, PWP2B, SWAP1, EFC12, NXRD1, ANR54, RM14, LTMD1, WDR93, EDC4, CP046, WDR73, PRS54, CP087, HAGHL, DCR1A, ATAD2, SPG17, ATRAP, ZN474, PLPR3, NHS, CC171, MAP12, GLB1L, BI2L2, RBP10, NSMF, CSC2A, DNMBP, DJC14, THADA, ZN497, LRC69, TMM91, DG2L6, PKHG7, MMS22, RHG27, FA47E, TM1L2, TTL10, KCP, UBP43, SMIM5, MED25, ZN765, CENPQ, MCM10, STRAA, MICA3, ZN367, APTX, TMC4, MUC19, DPOLN, SAXO4, CEA19, ADAT2, CATIP, ZN605, MYPN, YRDC, NHRF4, NLRX1, CAPS2, ST32C, THOC4, SESD1, WDR5B, TMM17, DAA11, NUD13, CNDG2, CTSR3, TXND6, DZIP3, PTH, TDRP, MYLK4, CASC2, RN135, CF337, ZN584, FMNL3, MSTN1, T151B, TRI59, CD302, CMA1C, DRC2, PGKA1, DRC7, CKP2L, DI3L2, SUV3, CCD13, SYTL1, REM2, CC170, KATL2, CFA46, RN168, DZI1L, AGRF2, CDK20, ASPM, TENS4, CENPL, ATOH7, F177A, ORML3, AHI1, CF107, ARG28, PYRD2, ABD12, LRC43, OR2LD, BBS5, PHTF2, TXND3, SVOPL, MINK1, ARL14, SPA6L, CCD24, LRC71, ARRD1, CC022, CCD33, TM148, CN178, BEND7, ADGB, CL076, ENDOV, S2529, CB069, IGS22, MUT7, CI163, SR1IP, ANR53, DAAF3, NLS1, NETO2, ZN485, MACA2, ADAD2, MORN4, CFA43, ZN248, TT21A, NSUN7, CS018, TTC16, DEN6B, ARMC2, DAAF1, TRPV1, WDR19, ANKK1, BHE22, BBS1, CC148, BEST4, NLGNY, CADM4, NEK11, OR1K1, O13C5, SVIP, DNJB8, CLHC1, GA2L2, ZSC18, PMFBP, ADA32, MIEAP, FANK1, SPP2B, SACA4, DLG5, NSUN6, TBCK, ZDHC1, NEUFC, S38A5, BLNK, OSCP1, SNR27, SKA2, TFB1M, STAG1, CQ050, TEKT4, DYDC1, SPIR2, S13A3, MB3L1, SREK1, MT21A, ASB13, ATS14, DYH7, PPM1E, TM234, CV015, TCPW, GBF1, LAR4B, NPIB3, PXDN, RSLAA, N42L2, DNSL2, OSTF1, HPS1, CLP1, WNT8B, SCAM4, TF3C6, LSM10, PMEPA, AKTS1, LENG9, ELMO3, ARL8A, ZN669, TIFA, EAF2, RB39B, DYH11, DRC10, RRFM, NGBR, OMA1, ZN837, CHFR, ADAT3, NEIL1, KIF12, IFT43, LRC46, HMCES, MED8, FAH2A, MSD3, CCD34, ZN845, RSRC1, TXD15, MRP9, MRP8, CHAP1, LRIQ1, BT3L4, DNJC1, RPGR1, ALPK3, IFT74, CCD17, CC74B, LRGUK, CFA53, KAD8, STAC3, HEXI2, PPM1M, CFA57, NOL4L, F218A, SPT33, CFA54, ZN396, SPT13, ODAD4, PCFT, AGAP4, FCRL4, ERMAP, ALS2, PRAM, ALPK1, 1A1L1, FCRL5, S26A8, MED15, IF140, CEP89, MBOA4, MMS19, INSM2, TBX5, TEAD3, CDCA3, SPSB2, RAD9A, ZN184, MGLL, ATX2, BARD1, FOXB1, SPAG8, RNZ2, TCF25, MEP50, LBHD1, PSD2, NEUL2, PIGQ, CPPED, HDHD3, GCP2, FEM1A, PHAF1, FUZZY, CHCH7, DERL1, DHRS6, RSRP1, KTNB1, KIFC3, DPCD, SMIM2, IFT27, THIC, NADAP, TM2D2, BBS2, TEKT3, IDI2, NUSAP, CAYP2, STK33, TRI55, BCDO2, DB126, FRMD8, PITM2, OSBL7, ULBP1, CHIA, B2L14, TSG10, SPEF2, ZFHX2, CFA74, ZN436, DYH6, LRC27, BSSP4, DNAI2, CHST5, CLPB, DRC9, ENKD1, TKTL2, MMP28, MIXL1, ZN106, VCX1, ABHGB, TOR3A, FA83D, COE3, MROH8, CSTP1, TM231, TASP1, CS044, CJ095, TBA4B, WDR76, CN093, COG4, MLXIP, KIF9, S28A3, SIAE, CLSPN, CYBP, NMUR1, IF122, PARVG, SPC25, LST2, ARGAL, NCK5L, LRC4C, SPCS, NRX3B, EMC7, UBE2T, SLAF7, TMM9B, TF7L2, IFT46, XPP3, ANM8, AAAS, STK36, SOSSC, GLRX2, CLTR2, KIF15, SEM4G, CT096, MITOS, KLH11, HAUS2, MED9, CB042, SIR5, MKS1, P4HTM, ARH38, PP4R2, BTG4, APC11, WSB2, MYOF, C1RL, CC167, KCMF1, RN181, SPAT7, CAC1I, DYH1, KIF17, C2D2A, MYEF2, ANKY1, SAE1, IF172, DPOLL, NAT8, RPGFL, NDUAC, SCNBA, DNAI1, TEKT2, PS1C1, KAD3, APC5, GGA1, DJC12, FBXL2, NXT1, RALY, TSN16, FBX5, INTU, TPC1, ABT1, AKP8L, GUC1B, PCDGG, PCDA6, RHG26, RBL2B, B9D1, ANR26, CE164, MOK, SPG7, LAMP3, COLQ, DLEC1, PSF2, KCNK6, INVS, ASF1A, FBW1A, VP9D1, SL9A8, WDR37, GSTK1, RT28, RPC10, UB2J1, COQ4, EXOS1, CBY1, DYR1B, AAKB1, NRX3A, C170B, SPEF1, IFT25, SHAN1, ASB4, CEP83, PCDGM, TM14A, CABIN, USH1C, TNR11, SCIN
Species: Homo sapiens
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Song J, Liu C, Wang X, Xu B, Liu X, Li Y, Xia J, Li Y, Zhang C, Li D, Sun H. O-GlcNAcylation Quantification of Certain Protein by the Proximity Ligation Assay and Clostridium perfringen OGA(D298N)(CpOGA(D298N)). ACS chemical biology 2021 16(6) 34105348
Abstract:
O-GlcNAcylation is an O-linked β-N-acetyl-glucosamine (O-GlcNAc)-monosaccharide modification of serine or threonine in proteins that plays a vital role in many critical cellular processes. Owing to its low molecular weight, uncharged property, and difficulty in distinguishing from β-N-acetyl-galactosamine (GalNAc), the lack of high specificity and avidity tools and sophisticated quantification methods have always been the bottleneck in analyzing O-GlcNAc functions. Here, we compared glycan array data of the mutant of Clostridium perfringen OGA (CpOGAD298N), O-GlcNAc antibody CTD110.6, and several lectins. We found that CpOGAD298N can effectively distinguish GlcNAc from GalNAc. Glycan array analysis and isothermal titration calorimetry (ITC) show that CpOGAD298N has a GlcNAc specific binding characteristic. CpOGAD298N could be used in far-western, flow cytometry analysis, and confocal imaging to demonstrate the existence of O-GlcNAc proteins. Using the CpOGAD298N affinity column, we identified 84 highly confident O-GlcNAc modified peptides from 82 proteins in the MCF-7 cell line and 33 highly confident peptides in 33 proteins from mouse liver tissue; most of them are novel O-GlcNAc proteins and could not bind with wheat germ agglutinin (WGA). Besides being used as a facile enrichment tool, a combination of CpOGAD298N with the proximity ligation assay (PLA) is successfully used to quantify O-GlcNAc modified histone H2B, which is as low as femtomoles in MCF-7 cell lysate. These results suggest that CpOGAD298N is a specific tool for detection (far-western, flow cytometry analysis, and confocal imaging) and enrichment of O-GlcNAcylated proteins and peptides, and the CpOGAD298N-PLA method is useful for quantifying certain O-GlcNAc protein.
O-GlcNAc proteins:
FSIP2, DYTN, P121C, ANM5, RGRF2, MSI1H, SNX2, IDHC, BAZ1A, AT10B, DDAH2, ADH1, ASSY, APOB, PPBT, SRC, K1C18, INVO, TRY1, A1AT1, O5AL1, PYC, ITPR1, GLU2B, ASSY, SON, DRD4, H2B1O, CP2DA, CXCR1, DHE3, MAP4, TRY3, BASI, LMNA, NU153, BRCA2, H2B1D, NAL11, CH60, ACTC, PI42B, HRH2, FKBP4, ANK3, ATM, UBP2L, ADRM1, TEN4, PAN3, FSIP2, TGO1, COA6, FRY, MYOME, SHE, HCFC1, ZN148, Q63800, SBP2, DYH17, A2MG, IQCE, AEDO, FBX38, QSOX2, NRAP, CNDG2, THIM, LONM, CLC10, MFS6L, DSEL, HMCS1, TACO1, CNTRB, NUP93, S2541, KCD17, MB213, CFA43, STAB1, SMRC2, DTX3L, GANC, GRK7, TRIM7, F227B, ADAD1, ADCYA, ERP44, FANCJ, TMM59, TBB2B, SPIT4, AP2B1, NUD12, I2BPL, CARD9, PABP3, ABHGB, WDCP, TDRD3, SUCHY, DCA13, KI21B, HEBP1, CTNL1, UBQL2, CAD22, TNIK, WDR3, RBM7
Species: Rattus sp, Homo sapiens, Mus musculus
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