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Xu S, Yin K, Xu X, Fu L, Wu R. O-GlcNAcylation reduces proteome solubility and regulates the formation of biomolecular condensates in human cells. Nature communications 2025 16(1) 40307207
Abstract:
O-GlcNAcylation plays critical roles in the regulation of protein functions and cellular activities, including protein interactions with other macromolecules. While the formation of biomolecular condensates (or biocondensates) regulated by O-GlcNAcylation in a few individual proteins has been reported, systematic investigation of O-GlcNAcylation on the regulation of biocondensate formation remains to be explored. Here we systematically study the roles of O-GlcNAcylation in regulating protein solubility and its impacts on RNA-protein condensates using mass spectrometry-based chemoproteomics. Unexpectedly, we observe a system-wide decrease in the solubility of proteins modified by O-GlcNAcylation, with glycoproteins involved in focal adhesion and actin binding exhibiting the most significant decrease. Furthermore, O-GlcNAcylation sites located in disordered regions and with fewer acidic and aromatic residues nearby are related to a greater drop in protein solubility. Additionally, we discover that a specific group of O-GlcNAcylation events promotes the dissociation of RNA-protein condensates under heat stress, while some enhance the formation of RNA-protein condensates during the recovery phase. Using site mutagenesis, inhibition of O-GlcNAc transferase, and fluorescence microscopy, we validate that O-GlcNAcylation regulates the formation of biocondensates for YTHDF3 and NUFIP2. This work advances our understanding of the functions of protein O-GlcNAcylation and its roles in the formation of biomolecular condensates.
O-GlcNAc proteins:
MEX3A, CNOT1, RGPD3, P121C, MYO1G, H7C0N4, PDLI1, CBX4, TAF4, RFXAP, PODXL, ARI1A, UTY, ABLM1, DVL1, KMT2D, HGS, MYPT1, XPO1, SET1A, PUR4, FYB1, PPM1G, EIF3H, IPO8, NUP42, KDM6A, ZZEF1, TET3, SI1L1, CP110, SYNJ1, IF4G3, TGON2, WIPF1, HTSF1, MYPT2, SMCA5, GANP, MA1A2, DIAP1, PLIN3, MAFK, KIN17, CUTA, N4BP1, CLAP2, BRE1B, PP6R2, ANR17, ZN217, FLNB, NCOR1, SPAG7, TACC1, GGYF1, ERLN1, WBP4, U520, WDHD1, RNH2A, STAM2, CBPD, STK10, TOX4, ST65G, TOX, PCF11, ENDD1, UBR5, AGFG2, KIF4A, MPZL1, IPO7, BAG4, SC24B, HERC2, ZMYM6, BAG3, CLPT1, ABL1, ADA, LDLR, HLAB, TFR1, CATA, GCR, KITH, HLAA, RPN2, ITB2, RLA2, RLA0, ITB1, RB, TPM3, SAP, LAMB1, SP1, PTPRC, ITA5, VIME, SNRPA, ROA1, ATX1L, DERPC, MYBA, CH60, PABP1, TPR, SKI, XRCC6, LAMP2, ITA4, HCLS1, KPYM, PO2F1, RAC2, IGLL1, RS2, TFE2, LEUK, CREB1, GCFC2, ATF7, SON, ATF1, TFE3, NFKB1, AL5AP, PUR2, ROA2, CBL, IF4B, AT2B4, APC, MGAT1, ARNT, DCK, PYR1, CALR, MAP4, CALX, NDUS1, PML, PDIA3, HMOX2, AMRP, CLIP1, ZEP2, HNRH1, ELF1, ICAM3, TTK, CAH8, BMI1, PTN7, RFC1, ADDA, NU214, MP2K2, HPCL1, VATA, CUX1, TXLNA, MLH1, PEX19, S19A1, LAP2A, LAP2B, STAT1, EPS15, MATR3, KI67, CRKL, UTRN, CAPZB, RFX5, PRC2A, NASP, FAS, CDK8, YLPM1, NU153, RBP2, TAF6, EMD, TCPQ, PAPOA, PLCD1, HCFC1, AFF3, ROA3, AGFG1, RBM5, ACLY, SMTN, RD23A, AF10, DSRAD, CADH4, SEC13, IF6, STX17, CORO7, NU107, SELB, TPIS, RAB2A, DCAF7, TIM10, CNBP, TBA1B, NOMO3, F193A, IGBP1, IF4G2, GTF2I, PHC1, NUCB2, TMF1, RT09, SARNP, FOXK1, FBLN2, FAM3A, RBM10, CDK6, HNRPU, SPTB2, FOXK2, RUNX1, CAP1, MEF2A, SP2, KMT2A, IF4G1, TLE1, TLE2, TLE3, TLE4, REL, GFPT1, GABPA, RING1, VAC14, EP300, AHNK, BRME1, MGAT2, GALT2, TROAP, TROP, BPTF, SFSWA, DPYD, TP53B, IRAG2, NFAC3, ROA0, AIMP2, GPS2, PABP4, S39A6, PICAL, HDAC1, STIM1, MTMR2, CUL4B, ASPP2, PEBB, NFYC, LRP8, VEZF1, BOP1, SEPT6, UBP2L, MLEC, ZMYM3, SRC8, LAGE3, PUM1, EPN4, RRP1B, NCOA6, GSE1, MEF2D, NFIX, ZN638, NUMA1, CND2, CND1, RB3GP, NECT1, PCBP1, PCBP2, ELOC, KS6A1, SC23B, JHD2C, ELF2, TAB1, NCOA1, ZYX, ADRM1, DPYL2, TAF9, DREB, DGUOK, QSER1, RHG31, AAK1, QRIC1, LRRF1, GNPTA, C2CD3, EPC2, PDCD4, BOREA, PAN3, BRD10, TGO1, PRC2B, PCID2, P4R3B, DCA10, SNP47, MANEA, ZN362, LRIF1, CAF17, CAMP1, UBAP2, CPTP, RC3H1, AHDC1, VPS53, RPRD2, ZN318, TASO2, ZMYM4, PAPD7, RN123, MAP1S, ARID2, NIPBL, LIN54, ANR54, PPR18, EDC4, SCYL2, GOLM2, NFRKB, RSBNL, MDEAS, BRAT1, ZC3HE, LARP1, FIP1, CRTC3, SHSA2, PI16, MCAF1, BCOR, GGYF2, NFXL1, QSOX2, YJ005, UBN2, MTSS2, NOL8, SSH2, ASXL2, SPT6H, LRC8D, EIF3M, MEPCE, KDM3B, TRM1L, ZCCHV, EFL1, KANL1, POGZ, ZFY16, NUFP2, MAVS, EMSY, I2BP2, HUWE1, YTHD3, KAISO, LDB1, LYRIC, RETR3, TSYL5, DZIP3, CACL1, P66A, C2CD5, HIPK1, ERI1, DEN1C, VRK3, ARI3B, MGAP, Z3H7A, CDAN1, ANKH1, SUGP1, GLT12, K2013, PHAR4, DCP1B, XRN1, RPTOR, NUP93, ZN687, FNBP4, NOPC1, ZBTB2, EMC1, CB069, EIF1A, KRI1, AFG2A, NHLC2, AVL9, XXLT1, GOLM1, SUMF2, TXND5, KCT2, CHSTE, SLAI1, TNR6A, PHC3, SP20H, KMT2C, BD1L1, ARI1B, NUP35, KNL1, MSPD2, NEDD1, GABP2, PIGX, STT3B, ALMS1, NEK9, NEK7, PO210, GEMI5, PIGO, MZB1, RAB2B, TM263, NUDT8, ZFN2B, HNRLL, PCNP, LMO7, ATX2L, P66B, GBF1, SMG7, RTF1, TOPB1, TSC1, PRCC, RREB1, CBP, DDX17, SEM4D, GSLG1, RENT1, GPKOW, FUBP2, ARHG2, UBP7, LPP, IGSF8, MRTFA, ACD, RAB37, PF21A, DOC10, DCPS, FAF2, TMCO6, ESS2, RMI2, TONSL, PDLI5, ZN414, VCIP1, PDLI2, ANCHR, ZFR, EP400, PRRC1, RBM14, QKI, PURB, SPAG5, CIC, MED15, ERGI2, ERBIN, LYSM1, MYADM, SEBP2, RUFY1, MINT, HTF4, CNN2, AGAP2, CSN8, CGRE1, DOK1, CTIP, SMAD5, S29A1, ARI3A, ANM1, DPH2, PELO, ERP44, ESYT1, CNPY3, WAC, MMTA2, HNRL1, ALG8, TMUB1, YJU2, PAPOG, SSBP3, JAM3, OSB11, NAA15, EIF2A, MK67I, YTHD1, KPCD2, UCK2, FTO, AMRA1, CEP44, BC11B, YIPF3, BRD8, XYLT2, I2BPL, MEGF9, EPC1, SLK, ADNP, ZN106, TM245, TMX1, CENPH, WNK1, ZHX3, DOCK5, PKHG2, KANL2, PKHA5, CLSPN, MYG1, RC3H2, LY9, TAF9B, S38AA, TM165, NCOA5, TANC2, TNR6C, CHD8, RESF1, APMAP, ARFG3, UBN1, GP108, INCE, ANLN, GEPH, BMP2K, RBM12, STAU2, INT10, GPT2L, CARF, PP4R2, TAB2, RM39, EURL, CDK12, HPBP1, ITSN2, ARHGC, CHMP5, FOXJ2, SLAI2, CT2NL, TBC14, RBM27, KANL3, FRM4A, ANFY1, GMPR2, ZN219, SE1L1, B4GT7, STK39, SEC63, SUN2, SRP68, SEPT9, UBQL2, S30BP, PFD2, NRBP, EI2BD, BTNL2, CDC23, GGA2, GMEB2, DP13A, MA1B1, ACINU, NUP50, MRTFB, ZBT21, YETS2, HECD1, TPX2, MGT4A, SYNRG, MINP1, SCAF8, SET1B, TBD2B, DICER, SRRM2, SMC3, SCML2, ZN148, CLC11, POLH, PLAP, TPPC8, WBP11, GIT1, ZN281, SIT1, HBS1L, SALL2, TRRAP, ARIP4, HECD4, CRBG1, HYOU1, ATG4B, KLF12, PRC2C, RBM7, CD2AP, SRPRB, GMEB1, TACC3, STK24, NCOA3, M3K4, ZHX2, NS1BP
Species: Homo sapiens
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Li G, Meng F, Zhong X, Yu K, Zhang N, Zhang K, Huang H, Li W, Zhang J, Wang W, Ren Y, Liu Y. DNA probe pulldown screening uncovers O-GlcNAcylation modulation of transcription factor DNA interactions. Scientific reports 2025 15(1) 40596516
Abstract:
O-linked β-N-acetylglucosamine (O-GlcNAc), a critical post-translational modification predominantly found in the nucleus, plays a substantial role in regulating gene expression by modulating transcription factors (TFs) activity. However, quantitative analysis investigating the influence of O-GlcNAcylation on protein-DNA interactions at a proteome scale remains undone. Herein, a pulldown screening approach using a consensus TF response element (catTFRE) was employed to unravel the impact of fluctuating levels of O-GlcNAcylation on the DNA binding efficiency of endogenous TFs/co-factors. Utilizing quantitative proteomics, we identified a substantial enhancement in the binding capacity of 241 nuclear proteins (NPs) to DNA sequences due to elevated levels of O-GlcNAcylation, whereas a decrease in DNA binding was observed for 2 NPs concurrently. Intriguingly, the O-GlcNAcylation elevation significantly enhanced the binding of 146 TFs/co-factors to specific DNA sequences. We further established that the O-GlcNAcylation of several Forkhead family TFs, including FOXA1 and FOXC1, notably enhances their binding to specific DNA sequences in living cells. Our research presents an efficacious approach to assessing the impact of O-GlcNAcylation on the interactions between proteins and DNA. This significantly enhances our understanding of the role O-GlcNAcylation plays in the regulation of transcription.
O-GlcNAc proteins:
RBM47, ESYT2, HACL2, VWA8, CNOT1, PGP, SMHD1, U119B, MET15, MYO1G, ADAS, PDLI1, BACH, HAX1, SPT5H, TAF4, CLIC1, BMAL1, SAP18, DNM1L, PLOD2, PSDE, BIN1, NOP56, DDX3X, TRI38, NFIB, PPP6, CDKA1, ABLM1, ADA10, CHK1, PSA7, TX1B3, TIM23, MYPT1, S27A2, XPO1, SYN3, ZN609, SR140, SET1A, ZN185, ATX7, OGT1, EIF3D, EIF3H, TNC18, MOT4, P4HA2, NUP42, ARPC5, CLOCK, MAFG, KDM6A, DHX15, RRP8, PRP4, SERA, PHF1, DC1L2, PSMD3, SNUT1, MCA3, HNRPR, PRPF3, TPD54, EMC8, MGA, KLF4, TIM44, PLRG1, ZN207, BUB3, ACTN4, BUD23, ENSA, SPOP, LANC1, AKAP8, CALU, ORC5, ZNRD2, MED14, NPA1P, ZC3H1, AQR, GANP, KDM1A, ACSL4, VINEX, CCNT1, PLOD3, TSN3, PLIN3, MAFK, UGDH, PQBP1, TBL1X, NBN, MITF, CPNE3, ZC11A, ANR17, H2AY, FLNB, NCOR1, SPAG7, RM33, SC22B, PR40A, MPPB, KTNA1, PSIP1, ERLN1, NDUS3, SRS10, SF3B1, CSDE1, U520, NU155, CRTAP, S2512, EIF3G, SOAT2, CCNK, SPF27, SPF30, RL1D1, CLPX, SYUG, DFFB, SRP72, RECQ5, ATG12, TOX4, K0754, SRBS2, SUN1, ERLN2, PRP6, SC31A, UBR5, AGFG2, SCAF4, DUS14, SPN1, NDUB8, RTN3, EPN2, LC7L3, MBD4, VAPB, 6PGL, ZBT7A, IPO7, SLU7, CD2B2, PSMG1, SC24B, ZBT11, SNP29, OXSR1, HS74L, LSM8, AP2A1, WIZ, BAG2, AIFM1, BPNT1, SNAI1, DDAH2, TXD12, MBD3, TOM40, NAPSA, ACL6A, CCNE2, LDHA, NB5R3, PGK1, A1AT, FOS, RASH, LDLR, LMNA, ALBU, TFR1, TRFL, FRIH, NU3M, MIS, VTNC, CATA, GBA1, FUCO, ALDOA, CYTB, GCR, SODM, KITH, K2C1, G3P, HLAA, CPNS1, HSPB1, TYSY, RPN1, RPN2, GNAI2, AT1A1, AT1B1, A4, APOD, ALDH2, PERM, PPBT, IF2A, RLA2, RLA0, JUN, ITB1, K1C18, K2C8, GELS, ATPB, IF4E, ENOA, NPM, ITAV, GDN, ACBP, LDHB, CATD, ANXA2, TBB5, SAP, CATL1, SYEP, CATB, HNRPC, FUMH, TSP1, SP1, ANXA6, HS90B, SRPRA, ODPA, RU2B, RU17, ITA5, NFIC, VIME, K2C7, GNAI3, ANXA5, SNRPA, HMGB1, DLDH, ROA1, RU2A, LKHA4, CTF8, MNS60, RO60, MYBB, LYAG, HLAC, KAP0, CH60, CLUS, BIP, HSP7C, GTR1, ODPB, LAMP1, ACADM, TOP1, TOP2A, G6PD, MPRI, PRPS2, PABP1, PCNA, ADT3, FA5, IMDH2, TPR, KCRU, SKI, ACTN1, XRCC6, NR2C1, LAMP2, EF2, K1C10, K1C13, PDIA4, PLST, T2FB, MIF, FPPS, KPYM, ENPL, MYL6B, RSMB, PO2F1, PLAK, ALDR, RAC2, ERF3A, EZRI, ATF2, FOSL2, MCP, NDKA, GNS, RS2, DESP, MUC1, CREB1, MGMT, CD36, STMN1, YBOX3, ZFX, JUNB, UBF1, JUND, ATF7, PTN2, DDX5, ARF4, PTPRA, SON, NELFE, RCC1, ATF1, ATF6A, XRCC1, LFA3, NUCL, ELK1, TFE3, CSK22, NFKB1, TYPH, TBP, RAB6A, PSB1, LMNB1, HXA5, CSRP1, FLNA, VDAC1, SDHB, CD9, PIMT, FBRL, UBA1, NDKB, ROA2, RFX1, KAPCB, QCR2, ITA6, SFPQ, PPIB, MAOM, SP100, NFYA, SAHH, COF1, IF4B, GATA2, THIL, MCM3, RS12, BRD2, TYY1, ATPA, PSA1, PSA4, ITA3, DDX6, PTBP1, SYVC, KAD4, 1433T, VATL, ARNT, RFA1, APEX1, PYR1, CALR, MAP4, CALX, PSA5, PSB4, PEX2, NDUS1, TEAD1, GRN, T2EB, 3MG, TKT, SPB3, ECHM, KCY, PEBP1, PDIA3, 2AAA, CDC27, HMOX2, PURA2, AMRP, RPB2, SDHA, METK2, HXA11, HXC9, CPSM, 3HIDH, PUR9, HNRH3, HNRH1, 1433S, STIP1, L1CAM, PRDX2, P5CR1, ELF1, CGL, RL9, CSTF2, MCM4, MCM7, HSP74, TRY3, GPC1, CTNA1, BMI1, PCGF2, PHB1, RADI, RFC4, RFC1, MYH9, MYH10, COPB2, SOAT1, ADDB, BASI, FUS, NU214, DEK, GLRX1, K22E, MP2K2, VATE1, RL4, PGM1, SPB5, ODO2, SRP14, NUP62, TAGL2, SYUA, ETFB, RBMX, VATA, HSPA9, IF4A3, TXLNA, TCPZ, PBX2, MLH1, STAT3, MDHM, ECHA, IF2G, S19A1, ELK3, ACTY, LAP2A, LAP2B, LPPRC, THIM, ECE1, MUC18, MATR3, GPDM, SSRA, RANG, PRS6B, VDAC2, MP2K4, KI67, RAGP1, RECQ1, NOP2, CRKL, NEDD4, IQGA1, STT3A, CAPZB, RL29, UCRI, RFX5, COPD, PSMD8, PRC2A, HSP13, AGRE5, DPOD2, NR2C2, RPIA, LMAN1, FAS, CDK8, RM19, AL7A1, CENPF, SYAC, SYCC, SYSC, MCM2, ACADV, YLPM1, RBM25, CLK1, NU153, RBP2, GSK3A, GSK3B, TAF6, GUAA, DNLI3, MRE11, IDH3A, EMD, CPT1A, SERPH, F10A1, MXI1, MAP2, PPT1, TCPQ, TCPD, FXR1, FXR2, RAB5C, RAB7A, SMCA4, IDH3G, GALK1, MECP2, DHB4, SPHM, HDGF, ROA3, 6PGD, IMA1, GDS1, MP2K6, AGFG1, HNRPF, TF2AA, MSH6, CAZA1, CRIP2, NUP98, PLK1, ACLY, SUCA, COPA, PTTG, SC24C, TCP4, ICLN, HIRA, SYYC, UBP14, HSP72, AT1B3, RD23A, RD23B, KAD2, SNAA, IF5, RAD, TERA, DSRAD, TPD52, SEC13, PSA, HNRH2, EIF3B, SYMC, DPOE2, ATP5I, IF6, NU107, SESN2, FOXL2, EPIPL, TPIS, SC61B, ACTB, IF4A1, PRPS1, SRP54, RAB2A, ARP2, ACTZ, CSN2, ARF3, CH10, STXB1, B2MG, WDR5, HNRPK, RS16, SMD2, ERF1, RS4X, ACTA, RL23A, RS6, H4, GBB1, RL32, RL8, PPIA, RS27A, RL40, AP2B1, 1433Z, IF5A1, RACK1, ACTG, YBOX1, TPM4, EF1A1, ACTS, TBA1B, TBA4A, TBB4B, CSK21, HBB, HBA, CXAR, NOP14, RBM6, PITX1, IF4G2, GTF2I, GSTO1, PO6F2, PRKDC, BASP1, RT22, RT05, RT11, RT34, RT09, HMGN5, SARNP, LACTB, TXN4A, RL19, FOXK1, RBM10, EM55, TFAM, VIGLN, CDK5, CLH1, HNRPU, SPTB2, TIAR, SET, FOXK2, RUNX1, CAP1, TFAP4, EXOSX, XPC, EWS, MEF2A, SP2, ODO1, KIF23, DSG1, SP3, RL18A, FKBP4, PLOD1, TOP2B, M2OM, DYST, CREM, KMT2A, CENPC, LMNB2, UPAR, KIME, TAP1, IF4G1, K1C17, SSBP, SRS11, EF1A2, CALD1, KDSR, SUH, GABPA, GABP1, PRDX1, RL18, C1QBP, CKAP4, ZO1, KHDR1, SRSF4, PP2BA, DHX9, LG3BP, MFGM, CD47, SSRP1, SLFN5, RBBP4, NCBP1, EP300, AHNK, MGAT2, GALT2, BST2, MPPA, SIA4C, NU160, TBL3, TF3C1, ASPH, SNF5, BPTF, NFIA, CHD3, TP53B, IFRD2, AIMP1, ILF3, LMAN2, FOXC1, PP1R8, ECH1, PRDM2, CSN1, KLF10, NR1H3, TADBP, ROA0, PAK2, PSMD2, DDX10, GPS2, SRSF5, SRSF6, TIF1B, G3BP1, STF1, NMI, KCAB2, SKP2, PABP4, GRB10, MTA1, DC1I2, ORC2, SNTB2, PPIG, TCOF, ADAM9, TUSC3, SMAD4, PICAL, TBB3, ASAH1, HDAC1, SNW1, STIM1, CUL4A, CUL4B, FHL1, RUNX3, THOC5, SPTN1, NOG2, KLF5, RUNX2, PEBB, NFYC, CKAP5, SCRB2, DAG1, VEZF1, DSG2, KEAP1, MORC3, SAFB2, EIF3A, UBP2L, MLEC, ZMYM3, DCTN1, DYHC1, EI2BA, ELOA1, MAP7, SRC8, FLNC, CAPR1, NF2L1, HABP2, DHX8, ITPR3, PLSI, TRIPC, PUM1, MDC1, EPN4, SMC1A, RRP1B, RRP5, BMS1, GANAB, MBTP1, 2A5D, LBR, CBX2, KIF22, MEF2D, CHD4, LASP1, ARI5B, NFIX, ZN638, NOLC1, NUMA1, GAPD1, SPCS2, EMC2, MO4L2, ABRX2, CND1, SUZ12, U5S1, SYK, RRS1, IF4H, BRD3, WDR43, ACOX1, EBP, PLEC, PON2, NONO, PTPA, PWP2, RNPS1, RCN1, PCBP1, MTFR1, SF3B3, PUM3, DLGP5, KS6A1, SF3A2, SC23A, SF3A1, SURF1, MED22, TAF5, TAF7, MARE2, TSN, SF01, PCH2, MED1, JHD2C, TRIP6, MARE1, ELF2, NAB2, AAAT, USF2, VAS1, RB11B, EZH2, ZYX, SEPT7, ADRM1, TAF12, DDB1, SNPC1, CDC37, DPYL2, SYPL1, TAF9, SRSF7, DREB, NRF1, PCKGM, DDR2, LY6K, HNRL2, PDS5A, ZN800, P3H1, RA1L2, MA7D1, TBRG1, LARP7, ACSF3, TPRN, EPC2, DHB12, UBP39, SETMR, YIF1B, ZN326, NOM1, SH319, RRP12, TOIP1, TDIF2, NU188, HP1B3, ZN362, SYRM, ZC3HD, LRIF1, RM02, UBAP2, RBM26, RIF1, MRCKA, RPRD2, BRE1A, TASO2, KHDR2, AR6P4, ARID2, GL8D1, NR3L1, MSL1, ABLM2, GLYAT, TWF2, K0930, SN12L, SDE2, LIN54, ALKB2, PPR18, CAVN1, CDC73, PRP8, SCYL2, GOLM2, RHGBA, NFRKB, RSBNL, KCD18, NCEH1, MDEAS, ZC3HE, SAMD1, C1TM, FIP1, CSPG4, PLGT2, SBSN, CF089, LCN15, CREL2, LRIG3, BCOR, MPRIP, CD109, SRCAP, UBN2, IKIP, CENPU, LARP4, HAKAI, NOL8, ASXL2, SND1, DDX46, TM10C, EIF3M, MCM10, NDUF7, CXXC5, APTX, ZCCHV, KANL1, NUP54, POGZ, NUFP2, SZRD1, CLAP1, HEAT3, HDGR2, BRAP, EMSY, I2BP2, RBBP6, YTHD3, TAF8, TMED4, CENPV, PHLB2, MYPN, ACOT1, PABP2, LDB1, PB1, YRDC, LYRIC, KTN1, LUZP1, CAND1, THOC6, DDX42, SPAS2, CACL1, P66A, RIOX2, CRLF3, VRK3, NAV2, CCD50, MISP, CTL2, MGAP, SFPA2, SFTA1, PHF6, CHERP, SUGP1, SUGP2, CCAR1, BAP18, RM41, KLHL7, MILK2, DHX37, YAF2, SRRM1, ZZZ3, K2013, ELMD2, CORA1, PELP1, TTC5, MIER1, NDUF2, NUP93, ZN687, CMTR1, LRC47, INT1, ABD12, CARME, FBXL6, SRBD1, NKAP, RP25L, ATPF2, RM50, CPSF7, ARFG2, EFNMT, ARFG1, PAF1, ENAH, DAND5, KRI1, MLKL, XXLT1, GOLM1, SUMF2, SUMF1, PGLT1, DHB11, TXND5, SERB1, CHSTE, LS14A, FSAF1, RB15B, PHC3, DIK2A, POPD1, SRFB1, SP20H, VP37A, PCAT1, BD1L1, NUP35, KNL1, PLBL2, DDX55, THOC2, WDR36, KI18A, GABP2, NPL4, STT3B, SPP2B, BRX1, STK35, NEK7, GPX8, PO210, GEMI5, FBF1, ZN384, WIPF2, SETD7, SMAP2, TM263, NU133, PDC6I, CO040, NUDC2, TWST2, SCFD1, PCNP, SPRY4, TRUB1, LMO7, CTL1, CKAP2, ATX2L, PHIP, RT4I1, NELFB, PALLD, PYDC1, PSPC1, P66B, BBX, CTBL1, ELYS, S39A7, H1X, RT27, NR4A3, PHF3, NDRG1, HS105, ZN592, LAR4B, NU205, PRCC, TFG, TAF4B, RREB1, HDAC2, SC65, SYMPK, GGH, DDX17, RAD50, CELF1, GSLG1, GPKOW, SMRC1, SMRD2, RAB8B, FUBP2, TNPO1, ARHG2, GLMN, KAT5, UBP7, POP5, LSM10, LMBL2, NCLN, FUBP1, SOSB2, PBIP1, SH3K1, PF21A, PINX1, P5CR2, INT12, FWCH2, FAF2, RMD1, MSI2H, RRFM, MTEF3, THA11, SGF29, DTBP1, PTCD3, MMAB, MCRS1, CERS2, SMRD1, TOE1, DDX27, SYMM, P121A, PDLI5, ERO1A, INT4, FUBP3, SYNM, VSIG2, HPDL, PAWR, RBM41, RSRC1, THOC3, CHAP1, S35E1, CLP1L, Z512B, ZFR, EP400, CNO6L, PRRC1, ZN512, YTDC1, SPT33, ZN830, RPR1A, RBM14, NED4L, QKI, LENG8, TRNT1, PP1RA, DEPD7, ATAD5, TEFM, MED15, EFGM, PIGS, TM209, IWS1, SIN3A, RBM15, MINT, NIBA2, YMEL1, HTF4, CNN2, SEC62, CDC5L, PSMD1, RING2, PARK7, EYA3, LGMN, PLIN2, POP1, SCAFB, TEAD3, TTC1, PHB2, CSN8, HCD2, SMAD5, ROAA, S29A1, CPNE1, MYD88, PKP2, SH3G1, GDF15, ARP5L, ZFY21, CMS1, DPH2, TBA1C, BCL7B, MBB1A, COR1B, TIRR, FSP1, PELO, ERP44, TACO1, ESYT1, CCM2, CNPY3, DIDO1, NDC1, ZSA5A, PAXX, HNRL1, RPP25, RCAF1, TBB2B, TM109, NUP58, GNL3, RIOK2, TIM21, THIC, RBM4, MND1, NADAP, SSBP3, RFIP5, ORNT2, NAA15, SRRT, AP1M1, NUSAP, HDHD5, RTF2, PECR, PDIP3, CEP41, RM37, GTPB4, UACA, UCK2, TB182, LNP, CEP44, WDR33, CTDS1, MFF, RB33B, NAT10, SP130, BRD8, RGAP1, CSTFT, LMA2L, CK054, F234A, RBM38, TDIF1, ALX4, SIL1, APC1, TMX4, WDR13, TARA, EPC1, ADNP, IPYR2, RM46, PININ, TM245, FOXP1, CPVL, GHITM, NELFA, BORG4, UN45A, SMOC2, AMPB, ZHX3, EMRE, REEP4, TM231, SH24A, DOCK5, CH033, SDS3, ACAD9, CQ10B, GORS2, JUPI2, SFXN1, SPE39, UBP42, RPF1, RPA2, GBB4, BRF2, RISC, CYBP, TENS1, TAF9B, TM165, MRM3, NCOA5, TRI39, SDF2L, AT131, RM47, APMAP, ARFG3, VTA1, PLPL8, SYSM, OSGEP, DMAP1, UBN1, RTN4, KMT5A, INCE, PRD10, ANLN, PDLI7, DDX21, MBNL1, SIR7, AAAS, ABCBA, PNO1, MBIP1, RAD18, SLX9, BMP2K, PDS5B, ECHD1, MDN1, DECR2, STAU2, DDX28, T106B, SPS2L, TYDP1, SEP11, DJC11, HAUS2, RBM22, MED9, ARGL1, SLTM, RMND1, AKIP, OXSM, THG1, HPF1, NDUBB, OCAD1, MARH5, MIC19, SIR5, CWC25, TEX10, THUM1, ING3, CARF, NSE4A, AATF, DDX56, TE2IP, ABI2, HXC10, BCLF1, TMOD3, MTCH1, HPBP1, MAT2B, ITSN2, CHMP5, THYN1, VAPA, RCC2, SLAI2, BCCIP, FPRP, RRBP1, CPSF2, RBM27, KANL3, GRHPR, NXF1, DAXX, TRM7, VTI1B, ABCF2, SYWM, SEC63, NDRG3, SUN2, LIMA1, SEPT9, TRPS1, DPP2, BAP29, NB5R1, S30BP, MED13, KAD3, HACL1, S2513, DBNL, CDC23, GMEB2, RCOR1, ACINU, NUP50, ZHX1, CDV3, CNO11, INT6, MCTS1, ZBT21, YETS2, ZMYD8, TMCO1, EPDR1, MYO6, KMT2B, PRP19, G3BP2, PACN2, MAGD2, PPIE, DIM1, MINP1, MACF1, PHF8, LIMC1, XCT, SRRM2, PA2G4, SMC3, SCML2, ZN148, RUVB2, VDAC3, RT33, PLAK2, ICE1, DDX52, MAN1, TR150, WBP11, MED16, NOP58, ZN281, SYYM, COQ6, LSM2, PHOCN, ORN, RT23, HDGR3, STRAP, RTCB, NOC2L, CHTOP, VPP2, TLN1, TRRAP, CRBG1, HYOU1, LAS1L, SAM50, PRC2C, RBM7, INSL6, YTHD2, PAXB1, SP16H, UTP18, CD2AP, SRPRB, TSSC4, SNX5, NUBP2, HCFC2, LRRF2, NPTN, 5MP1, DC1L1, CABIN, COX11, SCC4, ZHX2, MORC2
Species: Homo sapiens
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Levine ZG, Fan C, Melicher MS, Orman M, Benjamin T, Walker S. O-GlcNAc Transferase Recognizes Protein Substrates Using an Asparagine Ladder in the Tetratricopeptide Repeat (TPR) Superhelix. Journal of the American Chemical Society 2018 140(10) 29485866
Abstract:
The essential mammalian enzyme O-GlcNAc Transferase (OGT) is uniquely responsible for transferring N-acetylglucosamine to over a thousand nuclear and cytoplasmic proteins, yet there is no known consensus sequence and it remains unclear how OGT recognizes its substrates. To address this question, we developed a protein microarray assay that chemoenzymatically labels de novo sites of glycosylation with biotin, allowing us to simultaneously assess OGT activity across >6000 human proteins. With this assay we examined the contribution to substrate selection of a conserved asparagine ladder within the lumen of OGT's superhelical tetratricopeptide repeat (TPR) domain. When five asparagines were mutated, OGT retained significant activity against short peptides, but showed limited limited glycosylation of protein substrates on the microarray. O-GlcNAcylation of protein substrates in cell extracts was also greatly attenuated. We conclude that OGT recognizes the majority of its substrates by binding them to the asparagine ladder in the TPR lumen proximal to the catalytic domain.
O-GlcNAc proteins:
E2F8, PRXD1, TSPY2, CGB1, ZN839, CENPX, PDLI1, WWP2, FOXN3, IMA4, DPYL4, ABLM1, MEIS2, HGS, GAK, PLS1, MAGB4, ARPC5, DC1L2, FGF16, WIPF1, FRS3, RNF13, ZN207, ENSA, SMCA5, CE104, OGA, AP180, PQBP1, E2F6, ADAP1, DNJC8, FLOT1, TPPP, SCEL, CELF2, RPGF3, CLD14, KLF8, BAG3, PAK4, IGHG3, MBP, NFIC, SNRPA, CEL3A, RU1C, IGLC1, MTAP2, ERG, DMD, IRF2, ENPL, FOSL2, F261, SDC1, ATF1, ATF3, ELK1, TFEB, RXRA, COMT, USF1, FGFR3, NR4A1, THAS, MARK3, ARNT, PSB4, RBMS1, FCERG, MPIP1, CSTF2, CD68, BMI1, COPB2, SOX5, RS19, ARL4A, ETV5, IPP2, STAT6, ABL2, KSYK, TFPI2, RFX5, RXRG, CDK8, GSK3B, TAF6, SPS1, DNLI3, DYN2, GALK1, AGFG1, SC24C, DLX1, HDAC4, CTBP2, KLF3, TIRAP, PPIA, CSK21, 3BP2, SSBP2, CDKL1, BORG5, TFAP4, ROR2, CACB1, CREM, MECOM, TLE4, FAK1, COEA1, MTG8, GABPA, GABP1, DERM, TLE5, DEMA, ASPH, PP1R8, LCP2, DUS4, MTAP, EI2BE, G3BP1, GRB10, MTA1, PPIL2, PICAL, DYR1A, FHL3, NCOA4, CAYP1, CIRBP, DGCR6, VGLL4, CAPR1, GRB7, CASL, MEF2D, LASP1, SPCS2, SEPT2, PCM1, SIX1, SF01, TRIP6, RHOH, TAB1, ADRM1, CCDC6, BATF, NFE2, MAR1, NRF1, SMTL2, LETM2, OD3L2, AX2R, AMOT, AKIR2, PRR30, LARGN, RGPS1, PRC2B, TEX30, SNP47, PHYD1, KANK4, EEIG1, CPTP, AHDC1, KHDR2, EF1DL, OLM2B, ANKS6, RHG17, CNDH2, GRHL2, HMBX1, JHY, PP4R4, JMJD6, SCYL2, GOLM2, RSBNL, PSRC1, DRIC1, KCD18, FIP1, SAS6, CLM9, CCBE1, DUSTY, E400N, TM1L2, UBP45, MARK2, KCTD9, MCM10, CC186, POGZ, FLIP1, TAF8, RM55, S1A7A, SYVN1, PHLB1, CASZ1, PIM3, GA2L3, DDX42, CEP57, SPAS2, Z280B, PDPN, KCC1D, TF2LX, LOXH1, ARI3B, APLF, CC28A, RHG12, SKA3, PHC2, HYCC2, CMIP, SMAP1, RUSD2, ABI1, SYCE1, RPTOR, BATF2, S35F6, KLH36, ANS4B, MARHA, ZSCA1, UXS1, NECP1, BBOF1, LIPB2, MISSL, PHC3, TMIE, NUP35, OR4D6, JOS2, WDR48, GABP2, DPTOR, ALAT2, BRSK1, GRP4, HELQ, BL1S5, BCAS4, LMX1A, SSH3, SH3R2, FBF1, DLRB2, HDAC7, STYX, GTSF1, NEUR4, CSRN3, AP2B, SHC3, RORB, STAM1, CELF1, ARHG1, CSN5, RBPMS, ZDH16, PMEPA, UTP4, FERM2, SOSB2, R51A1, CR025, LENG1, BIRC7, PBIR1, CC126, PF21B, NRBF2, EDC3, PDLI5, ANCHR, ZC3HA, PKNX2, EGLN2, MARK4, UB2E2, PRRC1, NOL4L, AP4AT, SCLT1, TOX2, QKI, PNMA5, YPEL2, NUDC1, RB40C, UBL7, CK2N2, OSBL9, PNKP, PLIN2, CDCA3, HCD2, SNAG, ARI3A, DPYL5, CG025, SOSB1, TCF25, COE4, MIC25, UTP23, ALKB7, RBM42, LRRC1, TRMO, MCRI2, RSRP1, BBLN, ASHWN, SARG, NADAP, SSBP3, PLVAP, OSB11, B2L13, STK31, DGC6L, RM01, KLC2, SIK2, REN3A, NELFA, PPDPF, ILRUN, ES8L2, KI3X1, SP14L, B2L12, PKHA3, PKHA1, TAF9B, ARNT2, SYTL2, CIP1, CASS4, HINT3, PRDM5, PAR11, MBNL1, BRWD1, KLC4, TB22B, MBNL3, NFIP2, ZSC32, TMOD3, UGGG2, CWC15, KANL3, RBPJL, DKK4, LIMD1, DPOLL, AFF4, UBQL2, PUF60, BAZ2B, GGA2, ING1, BORG2, AGO2, ZHX1, MUCEN, ANR50, PCDGK, POLI, LIMC1, SMAG1, BAIP2, RPC10, PPIL1, C2CD2, MTL5, TBL2, YTHD2, T10B, SNX14, RT18B, GMEB1, STON1, PCLO, ZHX2, S23IP
Species: Homo sapiens
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Chai Y, Du Y, Zhang S, Xiao J, Luo Z, He F, Huang K. MicroRNA-485-5p reduces O-GlcNAcylation of Bmi-1 and inhibits colorectal cancer proliferation. Experimental cell research 2018 368(1) 29680296
Abstract:
Emerging evidences showed that miRNAs are involved in the oncogenesis of many cancers. Here, miRNA microarray analysis was performed to screen the significant miRNAs involved in the progression of colorectal cancer (CRC), miR-485-5p was chosen for further study. We found that the expression of miR-485-5p was significantly lower in CRC specimens and cell lines. In addition, low expression level of miR-485-5p is correlated with tumor progression and poor survival in CRC patients. Based on in vitro and in vivo assays, we found that miR-485-5p significantly inhibits CRC proliferation. Moreover, our results showed that miR-485-5p inhibits cell proliferation by reducing Bmi-1 protein expression, which has been reported to control the proliferation of many cancers. Mechanistically, OGT is a direct target of miR-485-5p, and miR-485-5p could inhibit the O-GlcNAcylation level of Bmi-1 by OGT. Overall, these results suggested that as a tumor suppressor, miR-485-5p may regulate CRC cells proliferation, which could regulate the O-GlcNAcylation and the stability of Bmi-1 through targeting OGT. This may give insight into a novel mechanism and therapy of CRC growth.
O-GlcNAc proteins:
BMI1
Species: Homo sapiens
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Li Y, Wang L, Liu J, Zhang P, An M, Han C, Li Y, Guan X, Zhang K. O-GlcNAcylation modulates Bmi-1 protein stability and potential oncogenic function in prostate cancer. Oncogene 2017 36(45) 28714959
Abstract:
The Polycomb group transcriptional repressor Bmi-1 often overexpressed and participated in stem cells self-renewal and tumorigenesis initiating of prostate cancer. In this progression, Bmi-1 protein was regulated by transcription and post-translational modifications (PTMs). Nobly, the underlying PTMs regulation of Bmi-1 is poorly known. Here we use co-immunoprecipitation show that in C4-2 cell line, Bmi-1 directly interacted with OGT which is the only known enzyme catalyzed the O-GlcNAcylation in human. Furthermore, we identified that Ser255 is the site for Bmi-1 O-GlcNAcylation, and O-GlcNAcylation promoted Bmi-1 protein stability and its oncogenic activity. Finally, microarray analysis has characterized potential oncogenes associated pathway subject to repression via the OGT-Bmi-1 axis. Taken together, these results indicate that OGT-mediated O-GlcNAcylation at Ser255 stabilizes Bmi-1 and hence inhibits the TP53, PTEN and CDKN1A/CDKN2A pathway. The study not only uncovers a novel functional PTMs of Bmi-1 but also reveals a unique oncogenic role of O-GlcNAcylation in prostate cancer.
O-GlcNAc proteins:
BMI1
Species: Homo sapiens
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