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Fan Z, Li J, Liu T, Zhang Z, Qin W, Qian X. A new tandem enrichment strategy for the simultaneous profiling of O-GlcNAcylation and phosphorylation in RNA-binding proteome. The Analyst 2021 146(4) 33465208
Abstract:
RNA-protein interactions play important roles in almost every step of the lifetime of RNAs, such as RNA splicing, transporting, localization, translation and degradation. Post-translational modifications, such as O-GlcNAcylation and phosphorylation, and their "cross-talk" (OPCT) are essential to the activity and function regulation of RNA-binding proteins (RBPs). However, due to the extremely low abundance of O-GlcNAcylation and the lack of RBP-targeted enrichment strategies, large-scale simultaneous profiling of O-GlcNAcylation and phosphorylation on RBPs is still a challenging task. In the present study, we developed a tandem enrichment strategy combining metabolic labeling-based RNA tagging for selective purification of RBPs and HILIC-based enrichment for simultaneous O-GlcNAcylation and phosphorylation profiling. Benefiting from the sequence-independent RNA tagging by ethynyluridine (EU) labeling, 1115 RBPs binding to different types of RNAs were successfully enriched and identified by quantitative mass spectrometry (MS) analysis. Further HILIC enrichment on the tryptic-digested RBPs and MS analysis led to the first large-scale identification of O-GlcNAcylation and phosphorylation in the RNA-binding proteome, with 461 O-GlcNAc peptides corresponding to 300 RBPs and 671 phosphopeptides corresponding to 389 RBPs. Interestingly, ∼25% RBPs modified by two PTMs were found to be related to multiple metabolism pathways. This strategy has the advantage of high compatibility with MS and provides peptide-level evidence for the identification of O-GlcNAcylated RBPs. We expect it will support simultaneous mapping of O-GlcNAcylation and phosphorylation on RBPs and facilitate further elucidation of the crucial roles of OPCT in the function regulation of RBPs.
O-GlcNAc proteins:
NACAM, SAP18, PLOD2, NOP56, DDX3X, PLXB2, RRP8, SERA, PSMD3, MCA3, PRPF3, TPD54, TIM44, ACTN4, ACSL4, PLOD3, IF2P, ZC11A, SC22B, PR40A, MPPB, CSDE1, U520, NU155, EIF3G, SPF27, RL1D1, CLPX, RTN3, LC7L3, VAPB, SMC2, AP2A1, WIZ, BAG2, TOM40, ACL6A, EGFR, LMNA, TFR1, FRIH, RPN1, RPN2, ITB1, SYEP, HNRPC, SRPRA, VIME, GNAI3, ANXA5, LAMP1, ACADM, TOP1, TOP2A, PABP1, ADT3, TPR, EF2, PDIA4, FPPS, ENPL, ALDR, NDKA, RS2, UBF1, ARF4, NUCL, RAB6A, PSB1, FLNA, SDHB, UBA1, NDKB, ITA6, SFPQ, IF4B, AT2B4, THIL, RS12, PSA4, SYVC, 1433T, MAP4, PSA5, PSB4, NDUS1, ECHM, KCY, AMRP, SDHA, METK2, CPSM, PUR9, HNRH1, 1433S, STIP1, P5CR1, MCM4, HSP74, CTNA1, MYH9, DEK, RL4, SPB5, NUP62, RBMX, TCPZ, ECE1, PRS6B, KI67, RAGP1, ATRX, SYQ, LMAN1, NASP, FAS, AL7A1, SYSC, MCM2, ACADV, NU153, RBP2, DNLI3, MRE11, CPT1A, F10A1, TCPD, RAB7A, IDH3G, HCFC1, DHB4, HDGF, ROA3, 6PGD, NUP98, ACLY, TCP4, SYYC, UBP14, SNAA, IF5, TERA, DSRAD, TPD52, EIF3B, NU107, EPIPL, SC61B, SRP54, B2MG, SMD2, RL23A, YBOX1, NOP14, IF4G2, GTF2I, NUCB2, RT22, HMGN5, RBM10, TFAM, CLH1, SPTB2, SET, CAP1, EXOSX, EWS, ODO1, RL18A, NUCB1, M2OM, LMNB2, SRS11, CALD1, RL18, C1QBP, CKAP4, KHDR1, DHX9, GOGA2, SSRP1, AHNK, AIMP1, ILF3, SRSF5, SRSF6, TIF1B, TCOF, PICAL, SNW1, TRI29, EIF3A, MLEC, CAPR1, SMC1A, RRP1B, GANAB, NUMA1, U5S1, RRS1, ACOX1, PLEC, RNPS1, PUM3, RB11B, SEPT7, DDB1, CDC37, SRSF7, PCKGM, HNRL2, INF2, PDS5A, PREP, RRP12, TOIP1, HP1B3, RBM26, BRE1A, CDKAL, PRP8, ZC3HE, QSOX2, IKIP, TM10C, EIF3M, PABP2, KTN1, CAND1, THOC6, P66A, MISP, CCAR1, PELP1, NDUF2, RM50, PAF1, TXND5, TOIP2, THOC2, TM263, NU133, PDC6I, SCFD1, LMO7, ELYS, RT27, HS105, NU205, RAD50, SMRC1, TNPO1, FUBP1, P5CR2, PTCD3, DDX27, EFGM, IWS1, NIBA2, YMEL1, PSMD1, EIF3C, ROAA, CMS1, MBB1A, GNL3, PDIP3, PININ, ACAD9, SFXN1, CYBP, RM47, RTN4, DDX21, AAAS, CARF, AATF, BCLF1, MYOF, SYLC, NXF1, SEC63, LIMA1, SEPT9, KAD3, RCOR1, ACINU, TMCO1, PPIE, PA2G4, RUVB2, TR150, RT23, CHTOP, TLN1, HYOU1, SAM50, SP16H, UTP18, SRPRB
Species: Homo sapiens
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Xie X, Wu Q, Zhang K, Liu Y, Zhang N, Chen Q, Wang L, Li W, Zhang J, Liu Y. O-GlcNAc modification regulates MTA1 transcriptional activity during breast cancer cell genotoxic adaptation. Biochimica et biophysica acta. General subjects 2021 1865(8) 34019948
Abstract:
Chromatin modifier metastasis-associated protein 1 (MTA1), closely associated with tumor angiogenesis in breast cancer, plays an important role in gene expression and cancer cell behavior. Recently, an association between O-GlcNAc transferase (OGT) and MTA1 was identified by mass spectroscopy. However, the potential relationship between MTA1 and O-GlcNAc modification has not yet explored.
O-GlcNAc proteins:
MED19, SBNO1, PGP, DPRX, SMHD1, RGPD3, ZSWM8, P121C, VGLL3, MCRI1, CK098, KIF2A, DX39A, PDLI1, BACH, MYO1C, MYO1F, AIP, SMAP, AP3B1, PSD11, PSD12, PSMD9, SPT5H, TAF4, DFFA, CLIC1, EIF3F, MATN2, LORF2, IPO5, SAP18, IF2B3, DNM1L, RTCA, AGRIN, PLOD2, HMGN4, IMA4, BCL9, PESC, NOP56, DDX3X, PODXL, CCN1, IMA3, NFIB, MA2B1, PDXK, ARI1A, TXND9, G3PT, CTRO, COPE, AP3D1, PDCD5, NHRF1, TPP1, TCRG1, TNPO2, PSA7, GIPC1, HAT1, HGS, MYPT1, HNRDL, XPO1, ZN609, SPTN2, SC16A, PLXB2, SR140, SET1A, LSM1, NPC1, ARC1B, ARPC2, ARPC3, TIF1A, PGRC2, PFD6, NKRF, LAMA5, ZN185, CASC3, OGT1, PMM2, HMGB3, PPM1G, FANCA, EIF3D, EIF3H, HDAC3, IPO8, STX7, NUP42, CLOCK, KDM6A, DHX15, MCES, FLRT2, PRP4, SERA, DC1L2, PSMD3, RFOX2, PAPS1, ZW10, SNUT1, SPIT2, TGFI1, M3K7, MCA3, HNRPR, PRPF3, TPD54, COCH, IF4G3, PPIH, ARK72, E41L2, TGON2, S26A4, FOXO3, DENR, XPOT, PLRG1, RGS10, ZN207, GET3, BUB3, ACTN4, BUD23, HTSF1, AP1G1, SYNC, KRT86, CPSF5, LANC1, AKAP8, CALU, RAD21, DHX16, MED14, SMCA5, JIP4, ZC3H1, TRAK2, AQR, GANP, KDM1A, GSDME, EVI5, SNX3, OGA, HNRPQ, CCNT1, PLOD3, AP180, PLIN3, MAFK, IMA7, UGDH, CTND1, SNX2, USO1, CCD22, PQBP1, DKC1, IF2P, EDF1, DNJA2, BRD4, PFD1, NBN, MCE1, WDR1, CPNE3, BRE1B, ZC11A, CLU, T22D2, PP6R2, CNOT3, CREST, ANR17, ZPR1, DUS11, PDCD6, TBCA, VATG1, H2AY, FLNB, NCOR1, SPAG7, SC22B, DEAF1, PR40A, VP26A, SAP30, MED24, KTNA1, PSIP1, SRS10, BAF, SF3B1, CSDE1, PRKRA, MED6, NPM3, LYPA1, U520, NU155, WDHD1, CRTAP, EIF3G, STAM2, CCNK, PRAF3, PIAS1, PIAS2, SPF27, DNJC8, SPF30, MPDZ, GLRX3, KRT36, RL1D1, CIAO1, SRP72, DDAH1, RECQ4, MTA2, MYO1D, TOX4, SC24D, PHF14, UBXN7, SUN1, PLPHP, ERLN2, PRP6, PCF11, NFAT5, HAUS5, AP2A2, SC31A, HEXI1, SEM3D, UBR5, AGFG2, SCAF4, ZFPL1, EPN2, ZRAB2, LC7L3, KAT7, MPZL1, FKBP9, CELF2, PAPS2, SMC2, LYPA2, IPO7, AGM1, CD2B2, BAG4, AHSA1, PSMG1, SC24A, SC24B, AGRL2, ACSL3, PCNT, CNOT4, CPSF4, EYA4, K2C75, SNP29, OXSR1, HS74L, PTBP3, AP2A1, STAU1, CAVN2, BAG2, M4K4, BPNT1, MBD3, CLPT1, ACL6A, CYB5, ADH1B, ADH1G, LDHA, DHE3, NB5R3, GSHR, SODC, PNPH, HPRT, AATM, EGFR, PGK1, KAD1, LALBA, FA10, TRYP, ASSY, A2MG, LDLR, HLAH, HBA, CO1A1, K1C14, K2C6A, LMNA, CASA1, CASA2, CASB, CASK, FINC, LACB, ALBU, ALBU, TFR1, GLCM, ALDOA, CYTB, ANXA1, GCR, PRIO, KITH, K2C6B, K2C1, G3P, CPNS1, HSPB1, RPN1, RPN2, GNAI2, AT1B1, ALDOB, A4, ARGI1, S10A8, HMGN1, ISG15, IF2A, HMGN2, ICAM1, RLA1, RLA2, JUN, LA, AP2A, ITB1, K1C18, K2C8, MYL1, GELS, PTMA, CDK1, ATPB, S10A6, IF4E, ENOA, PYGL, G6PI, DPOLB, NPM, TPM3, ITAV, LIPL, HEXA, ACBP, FABPL, LDHB, PDIA1, H10, CATD, ANXA2, CAN1, TBB5, TRY1, SAP, HEXB, PROF1, APT, K2C4, SYEP, CATB, HS90A, HNRPC, LAMB1, TPM2, FUMH, TSP1, SP1, ANXA6, DAF, MDR1, 4F2, HS90B, ASNS, ADH4, HEM3, RU2B, MET, RU17, ITA5, NFIC, VIME, RS17, K2C7, K1C13, ANXA5, K1C16, RSSA, CD63, SNRPA, GSTP1, RU1C, LEG1, HMGB1, CLCA, ROA1, RU2A, PARP1, UCHL1, LKHA4, ALDOC, CC175, ATX1L, ZN865, CT45A, HS71B, CALM3, RO60, H14, PTPRF, THIO, PPGB, KAP0, HEM4, ESTD, CH60, BIP, LAMC1, PPAL, HSP7C, GTR1, PYGB, LAMP1, TOP1, TOP2A, G6PD, C1TC, MPRI, ADHX, SRF, PABP1, PCNA, HARS1, IMDH2, TPR, KCRB, ANXA3, ACTN1, XRCC6, XRCC5, LAMP2, RINI, MYH6, ICAM2, AT1A3, EF2, K1C10, K2C5, PDIA4, TCTP, PLST, GYS1, T2FB, CD59, MIF, GLU2B, FPPS, TRI27, AK1A1, KPYM, ENPL, CCNB1, PO2F1, HNRPL, SYDC, ETS1, PLAK, ALDR, AMPN, PVR, ERF3A, EZRI, ATF2, FOSL1, FOSL2, MCP, NQO1, GNS, ZEP1, RS2, TFE2, DESP, RFA2, CD44, ITB4, CBR1, CREB1, BGAL, H15, H13, H12, AT2A2, STMN1, YBOX3, HSP76, HMGA1, JUNB, UBF1, JUND, ATF7, CAN2, PTN2, PYRG1, DDX5, PFKAL, LEG3, PRS6A, TCPA, PTN1, ARF4, RL7, VINC, SON, NELFE, RL17, PGAM1, RCC1, ATF1, XRCC1, CADH2, ML12A, NUCL, HXK1, RO52, SPEE, SL9A1, CSK22, RXRA, NFKB1, IF2B, ANXA7, TBP, BTF3, PSB1, MPRD, LMNB1, ICAL, CD11B, VATC1, CSRP1, FLNA, 5NTD, TAF1, CD9, TGM2, PIMT, FBRL, PUR2, PUR6, UBA1, NDKB, ROA2, RFX1, CBL, EST1, FBLN1, TCEA1, ITA6, SFPQ, PPIB, SYWC, RS3, NFYA, SAHH, COF1, IF4B, KTHY, CCND1, COT2, EF1B, PPAC, RPB1, CDK2, MCM3, NFYB, THTM, LAMA1, RS12, BRD2, TYY1, DNJB1, ATPA, PSA1, PSA3, PSA4, ITA3, MOES, DDX6, CTNA2, DNMT1, PAX6, U2AF2, RL13, HMGB2, PTBP1, SYTC, SYVC, EF1G, 1433T, ARNT, RL10, RFA1, APEX1, PYR1, CALR, MAP4, CALX, PSA5, DPOD1, MK01, RXRB, GRN, PTPRM, T2EA, CCN2, EPHA2, 3MG, KDM5A, TKT, RBMS1, PML, EF1D, ERP29, PRDX6, BLVRB, PRDX5, RL12, KCY, PEBP1, PDIA3, 2AAA, 2AAB, NMT1, HMOX2, PURA2, UFO, PUR8, CLIP1, SORCN, ILEU, RPB2, SDHA, METK2, CPSM, TIA1, ZEP2, DNJA1, AKT1, PUR9, HNRH3, HNRH1, 1433B, 1433S, STIP1, S10AB, L1CAM, PRDX2, OTX1, CDD, ELF1, RL9, KINH, CSTF2, DUT, TTK, MCM4, MCM5, MCM7, HSP74, GPC1, PROF2, CTNA1, CTNB1, PCGF2, SPB6, RADI, RFA3, RFC4, RFC2, RFC1, RL22, T2FA, SPRE, HXD13, CBS, K1C9, FBN2, GDE, MYH9, MYH10, COPB2, BASI, FUS, NU214, DEK, K22E, RL4, PP1G, GNL1, SRBP1, SRP14, NUP62, TAGL2, TALDO, RBMX, COIL, VKGC, GRP75, IF4A3, RS19, RL3, COIA1, OST48, AN32A, FEN1, CAPG, IL6RB, TXLNA, TCPZ, NNMT, PBX2, RL13A, STAT3, MDHC, MDHM, RFC5, IF2G, CETN2, BUD31, NAA10, CSK, GARS, SYIC, EIF1, KPCI, LAP2A, LAP2B, STAT1, MTREX, RS27, HELZ, RL35, CDN2A, PCP, ECE1, LIS1, MUC18, MATR3, MSH2, SSRA, RANG, NAMPT, PRS6B, NKX21, VDAC2, CBX5, UBP5, KI67, RAGP1, RECQ1, NOP2, CRK, CRKL, BAG6, NSF, NOTC1, MP2K3, RL27A, RL5, RL21, RL28, RS9, RS5, RS10, MAP1B, GNPI1, YAP1, UTRN, IQGA1, STT3A, CAPZB, IF1AX, SYQ, RL29, LIMS1, PPCE, RFX2, RFX5, SOX9, COPD, GSH0, PSMD8, PRC2A, TCPE, NEST, HSP13, KC1A, PIPNB, AGRE5, DPOD2, PAXI, RL34, NASP, FAS, CDK8, TCPG, EFTU, SRP09, PCY1A, SYAC, SYCC, SYSC, PRI2, CTCF, PSB2, MCM2, TSP3, YLPM1, TMEDA, RBM25, NUMB, HINT1, NU153, RBP2, GSK3A, GSK3B, TAF6, SPS1, GUAA, DNLI3, MRE11, CRIP1, GDIB, EMD, SERPH, F10A1, VASP, DYN2, CDK9, BCAM, PPT1, RL14, TCPQ, TCPD, ANX11, PAPOA, FXR1, FXR2, RAB5C, RAB7A, DUS3, SMCA4, GALK1, SSRD, HCFC1, UBP11, NR0B1, HDGF, ROA3, 6PGD, HNRPM, IMA1, IMA5, GDIR1, AGFG1, HNRPF, TF2AA, MSH6, ZN131, ZN143, RBM5, SPSY, EFNB2, CAZA1, CRIP2, NUP98, PPP5, ACLY, MAP11, COPB, COPA, CATC, AP2S1, PTTG, SMTN, IST1, SC24C, TCP4, ICLN, SYRC, ATX1, ATN1, SYYC, UBP14, HSP72, BCAT1, AT1B3, RD23B, PRRX1, HMCS2, SNAA, IF5, DRG2, XPO2, TERA, MFAP1, AF10, AF17, NP1L1, ADK, DSRAD, LAMB2, FOXG1, TPD52, SEC13, NH2L1, PSA, HNRH2, SUMO3, EIF3B, GFRA1, SYMC, IF6, CTBP2, TMM33, PCBP3, PCBP4, CORO7, NU107, MTPN, H2B1D, DEF3, ARPC4, SC61G, TPIS, EIF3E, SC61B, PP4C, MYL6, ACTB, IF4A1, RS20, PRPS1, PSA6, S10AA, CDC42, DEST, GMFB, RAB8A, SPCS3, RAB2A, RAB5B, RAB10, UBC12, UBE2N, ARP3, ARP2, ACTZ, CSN2, ABCE1, RS3A, RL26, PSME3, RL15, RL27, RL37A, RHOA, S61A1, LYSC, COPZ1, SUMO2, DCAF7, WDR5, NTF2, HNRPK, 1433G, RS7, PP1A, PP1B, PRS4, PRS8, RS8, RS15A, RS16, UB2G1, 1433E, RS14, RS23, RS18, RS29, RS13, RS11, RUXE, RUXF, RUXG, LSM3, LSM6, SMD1, SMD2, SMD3, TYB4, PRS10, RL7A, RB11A, ERF1, CNBP, RS4X, RL23A, RS6, H4, H2B1C, RAB1A, RAN, RL23, RAP1A, UB2D2, RS24, RS25, RS26, RS28, RS30, GBB1, RBX1, GBB2, RL30, RL31, RL10A, RL32, RL11, RL8, PPIA, FKB1A, RS27A, GRB2, TRA2B, RAC1, AP2B1, 1433Z, RSMN, SUMO1, DYL1, RL38, GBG5, RS21, IF5A1, RACK1, ACTG, SPT4H, UBC9, TYB10, PP2AA, YBOX1, CSK2B, TPM4, UB2L3, EF1A1, ACTS, TBA4A, TBB4B, CSK21, PA1B2, HBB, HBA, F193A, IGBP1, RBM6, IF4G2, RPP30, GTF2I, CNTP1, PHC1, TCPB, KRT85, RAE1L, PRKDC, MT1X, NUCB2, DCD, SSBP2, SARNP, RL24, CBX1, ARF1, ERH, RL19, SRSF3, FOXK1, DAB2, FBLN2, PGBM, RHG04, EFNB1, RBM10, RBM3, 2ABB, PIPNA, MPCP, VIGLN, TF2B, CDK5, CLH1, HSF1, FKBP3, DHSO, HNRPU, U2AF1, SPTB2, TIAR, SET, SRSF2, FOXK2, RUNX1, FABP5, CAP1, NC2B, TFAP4, EXOSX, OTUD4, PFKAP, EWS, TAGL, AK17A, MEF2A, KIF23, NRG1, DSG1, SP4, SP3, RL18A, MP2K1, FKBP4, PLOD1, NUCB1, RL6, TOP2B, DYST, CREM, KMT2A, LMNB2, TF65, UBXN1, GLGB, IF4G1, K1C17, NOTC2, TLE2, TLE4, LGUL, AK1C1, 1433F, CSTF1, PTN12, FAK1, SRS11, EF1A2, CALD1, PTN11, GFPT1, EXOS9, PSME1, SUH, GABPA, GABP1, RING1, PAX8, FMR1, PRDX1, RL18, CKAP4, ZO1, KHDR1, BAX, KLC1, LRP1, SRSF1, SRSF4, DHX9, GOGA2, LG3BP, CD47, PPID, SSRP1, NSUN2, RBBP4, EP300, AHNK, APOBR, HSP7E, MGAT2, GALT2, AP1B1, CPSF1, BST2, WASC5, NU160, SCAP, SRBP2, TBL3, ASPH, TROAP, TROP, SNF5, BPTF, CDC20, FSTL1, GRSF1, SFSWA, CHD3, SF3A3, TP53B, AIMP1, ILF2, ILF3, LMAN2, ELAV2, FOXC1, PP1R8, CSTF3, FLII, COASY, ACACA, CSN1, CAF1B, RED, MTAP, TADBP, ROA0, AIMP2, FADD, PRDX4, PAK2, CBX3, STK3, PSMD2, GPS2, SRSF9, SRSF5, SRSF6, TIF1B, G3BP1, CLN3, PTK7, PABP4, IFIT5, MTA1, EIF3I, 2A5G, CTBP1, UB2V1, DC1I2, ILK, MSLN, PPIG, TCOF, SF3B2, HAP28, ADAM9, FKBP5, SMAD4, PICAL, SQSTM, TBB3, ASAH1, PRP4B, PIN1, RIPK1, HDAC1, KCC2G, DCTN2, SNW1, STIM1, TRA2A, PWP1, CUL1, CUL2, CUL3, CUL4B, ASPP2, DYR1A, TPBG, FHL1, PM2PB, MOGS, CD166, NACA, THOC5, SPTN1, PKP1, DX39B, EXOS2, TBB2A, KLF5, BYST, IDI1, RUNX2, PEBB, NFYC, CDK13, CKAP5, CIRBP, COTL1, HNRPD, SCRB2, NID2, DAG1, VEZF1, DSG2, SAFB2, EIF3A, UBP2L, SCRIB, MLEC, TTL12, FHL2, DCTN1, DYHC1, EI2BA, IF4A2, SRC8, TRI25, FLNC, FA50A, MYPC2, GAMT, CAPR1, RBM39, KT33B, HLTF, PDIA5, DHX8, MCM6, ITPR3, MDC1, EPN4, SMC1A, RRP1B, NCOA6, GSE1, PSF1, UBP10, GANAB, 2A5D, LBR, GOGB1, MEF2D, CHD4, LASP1, NC2A, ZN638, IMB1, NOLC1, NUMA1, GAPD1, K2C72, NAA25, FL2D, SEPT2, SART3, U5S1, S39AE, SETB1, IF4H, BRD3, EEA1, PDIA6, PA1B3, PLEC, NOMO1, IPYR, TEBP, NECT1, NONO, PTPA, RBBP5, RCN1, K1H1, PCBP1, PCBP2, ELOC, SF3B3, DLGP5, RSU1, CNN3, SAFB1, SF3B4, SF3A2, RBMS2, SC23A, SC23B, SF3A1, SSXT, TAF5, TEAD4, TGIF1, NCOA2, NHRF2, TRAM1, SF01, TRIPB, PCH2, MED1, HMGN3, JHD2C, TRIP6, MARE1, ELAV1, ELF2, TAB1, AAAT, MLF2, TOM34, TBCE, UB2V2, STXB2, NEDD8, USF2, VAS1, VPS72, ZFHX3, ZYX, SEPT7, ADRM1, CCDC6, UAP1, AINX, PSMD5, CSRP2, DDB1, CDC37, DPYL2, RBBP7, TAF9, SRSF7, CPSF6, SMN, NRF1, FSCN1, IF16, GLO2, H2B2E, CGT, TRXR1, LY6K, HNRL2, Q1RMN8, PDS5A, QSER1, AAK1, WASC4, PRSR3, TSR1, QRIC1, SMU1, P3H1, MA7D1, CHD9, LSM12, GELS, A1AG, TBB8, GLYR1, FIL1L, AMOT, FA98B, EPC2, RHG29, FND3B, CRTC2, DHB12, GLE1, SNUT2, LACB2, P5CR3, PKHG4, H90B2, R39L5, ZN326, FILA2, BCORL, FIGN, K2026, AG10B, CC038, TGO1, PRC2B, RRP12, TOIP1, PCID2, SPXN3, SMAG2, BRM1L, TM201, NU188, HP1B3, CE170, ZN362, EYS, FKB15, DDX59, ZC3HD, LRIF1, UBR4, UBAP2, KPRP, XP32, RBM26, NT5D1, AHDC1, RPRD2, PR38B, BRE1A, ZN318, BROX, TASO2, ECM29, MBNL2, ZMYM4, K2C79, ARID2, USF3, C19L1, VIR, K1C40, HEAT6, BICRL, ATLA3, CARL2, CPIN1, TWF2, P4R3A, NIPBL, PDE12, LIN54, TET2, TM214, ARMC6, CAVN1, ERAP2, CDC73, TAF2, MED27, EDC4, PRP8, SCYL2, TTC27, GOLM2, NFRKB, MZT2A, CTR9, NCEH1, TRI65, MDEAS, BRAT1, ZC3HE, LARP1, BRWD3, SMRD3, FIP1, CRTC1, SAS6, CSPG4, CWC27, WDR82, ATL4, MCAF1, PACS1, MPRIP, GGYF2, CD109, CERS6, QSOX2, SRCAP, RIPR1, YJ005, UBN2, FGD6, PAXI1, RAPH1, IKIP, H32, LARP4, MED25, TBA1A, H2AV, HAKAI, ASXL2, CCD80, BT2A1, SPT6H, SND1, DDX46, BZW1, RSRC2, CYFP1, KDM3B, OLIG3, PHF5A, ZCCHV, KANL1, NUP54, POGZ, MYH14, NUFP2, CLAP1, PLGT3, HEAT3, HDGR2, EMSY, RAI1, I2BP2, RBBP6, SH3R1, HUWE1, YTHD3, KHDC4, TMED4, CENPV, T179B, Q862S4, KAISO, PATL1, MYPN, PEG10, NOP9, PABP2, MTA70, LDB1, PB1, LYRIC, TLK2, ZN598, KTN1, BCL9L, DTX2, LUZP1, THOC4, GP180, PACS2, CAND1, TSYL5, THOC6, MET16, NIPA, CARM1, PRSR1, MEX3D, DDX42, DAAF5, P66A, HORN, RB6I2, RIOX2, RBM45, PLD3, NUDC3, AHNK2, FOXP4, TEX2, Z3H7A, PHF6, CHERP, ANKH1, SUGP1, CCAR1, SLF2, PHC2, BAP18, RB12B, RAVR1, SPB1, IHO1, SRRM1, SMAP1, P20D2, KATL2, PHAR4, TRM2A, DCP1B, NALCN, MAML2, PELP1, ABI1, PORIM, CCAR2, NUP93, ZN687, CMTR1, LRC47, K2C78, ABD12, PALS1, STAG2, FNBP4, TAB3, DPP10, CPSF7, ARFG1, EMC1, PAF1, UBR7, ENAH, KRI1, PR38A, F184A, KDM1B, NHLC2, GT251, PGLT1, DHB11, TXND5, PAIRB, LEMD2, FA98A, RN214, LS14A, MISSL, TNR6A, PHC3, ABCF1, NGDN, SP20H, VP37A, KMT2C, ZMIZ2, ARI1B, NUP43, NUP37, NUP35, STXB6, KNL1, PLBL2, RLA0L, THOC2, WDR36, SMRC2, NPL4, PUM2, RDH11, STT3B, SPP2A, HM13, ALMS1, C99L2, MK15, BRX1, RBCC1, DOT1L, PO210, GEMI5, PARD3, IPO4, ZN384, K2C79, NOC3L, SCRB1, SMAP2, CCD12, TM263, NU133, PDC6I, GATD1, THEM6, CTF18, NUDC2, SCFD1, HNRLL, UBCP1, PCNP, LMO7, ATX2L, PHIP, PRP31, NELFB, PALLD, SREK1, PSPC1, P66B, DYH7, DNJC9, BBX, CTBL1, KAT6B, ELYS, ZCH14, DDX1, H1X, PSMF1, SMG7, RTF1, NICA, TM131, ELMO1, BAP1, PHF3, MAML1, NDRG1, HS105, CNOT9, ZN592, LAR4B, GCN1, PRP16, NU205, AN32B, NECT2, PRCC, TFG, TAF4B, HDAC2, STAM1, REQU, CBP, KAT6A, SYMPK, RBP56, GGH, DDX17, RAD50, CELF1, OSTF1, UFD1, RENT1, CSN5, GPKOW, SMRC1, SMRD2, FUBP2, TNPO1, NEP1, GLMN, USP9X, UBP7, CUL5, STMN2, LPP, MED12, H2A1C, BHMT1, SMCE1, MYDGF, RL36L, PSMG2, NCLN, UTP4, PP14A, MGN2, FERM2, FUBP1, LRC59, FKB10, PF21A, PPWD1, INT12, FWCH2, KLH29, CC124, AP2M1, REPS1, SNR40, MBD6, RMXL1, SEH1, TCAL4, THA11, GNA1, T3HPD, DAZP1, SAAL1, SGF29, RBM33, TMA16, ZN503, DYL2, CPNE2, THOC1, OTUB1, HMCES, MED8, DUS3L, DCNL1, SMRD1, DDX27, GWL, LIN37, ZCCHL, P121A, PDLI5, ERO1A, C2AIL, MED30, INT4, FUBP3, SPF45, ABHEB, PAWR, THOC3, KIRR1, WNK4, COG3, TMX3, CHAP1, LCOR, PDLI2, ANCHR, ZC3HA, TOPK, SCYL1, EHMT2, ZFR, EP400, PRRC1, FXL18, CCD43, NOL4L, DACH2, NSUN5, RPR1A, IPO9, RBM14, QKI, ALS2, PP1RA, S38A2, VPS35, PURB, PHF12, XPO6, NACC1, CIC, MED15, MCCA, NUDC1, ERBIN, WRIP1, RANB9, MYADM, SRPK1, STRBP, TM209, IWS1, SIN3A, RSF1, RBM15, RUFY1, MINT, PNKP, MMS19, UHRF1, NIBA2, HTF4, TBCB, CNN2, CDC5L, PSMD1, PFD5, RING2, PARK7, EYA3, VAT1, LGMN, NUP88, POP1, MNT, SCAFB, EIF3C, TTC1, DNJC7, KIF2C, RGPD5, ATX2, ROAA, NP1L4, SNAG, TS101, CPNE1, TCPH, EBP2, ARI3A, ANM1, H2A1J, PKP2, SEM3C, VRK1, RBM4B, DDX50, RNZ2, MEP50, TBA1C, MBB1A, COR1B, TXD17, CPPED, PDD2L, PYGO2, LLPH, PSF3, ERP44, GCP2, ESYT1, SEN34, CSN4, WAC, DIDO1, RBM42, MCMBP, DCNL5, RAMAC, AN32E, NDC1, MMTA2, TBB6, HNRL1, PHF23, DDX23, MCRI2, WDR18, MTND, NTM1A, TBB2B, ASHWN, TM109, UT14A, TMED9, NUP58, SELO, DPCD, GNL3, KIFC1, NUP85, ELOV1, THIC, RBM4, GATA5, LS14B, RBM24, TRPM6, NAA15, SRRT, AP1M1, MAK16, ITPA, RTF2, EIF2A, PDIP3, TMPSD, MK67I, YTHD1, WNK3, GTPB4, UACA, CRNL1, TBL1R, PP12C, API5, DPY30, TRIM4, GRB1L, UNK, TB182, AMRA1, TANC1, TTYH3, WDR33, TINAL, NDEL1, PIP30, MKRN2, ILKAP, XRN2, TOLIP, SP130, RGAP1, CSTFT, RAB1B, LMA2L, ING2, I2BPL, TMX4, EPN3, MED28, SH3L3, PPIL3, ADNP, DHX36, PININ, PABP3, CPVL, BOLA2, TMX1, NELFA, PTN23, UN45A, CHM4B, OFUT1, WNK1, AMPB, ZHX3, EHD1, HEAT1, SUV92, CCD86, PRR36, BCAS3, RANB3, PEAK1, SMYD3, WDR26, SDS3, SFR19, KAT8, PHAX, UBE2Z, RMD5A, BRD9, GORS2, JUPI2, GHC1, MED20, CNO10, KANL2, FBRS, SP30L, MLXIP, XPO5, MYG1, CYBP, RC3H2, TENS1, SPC25, TAF9B, GLOD4, NCOA5, MOV10, TNR6C, CHD8, SYF1, SRA1, CHM1A, TM9S3, APMAP, ARFG3, HM20A, SSU72, VTA1, ENY2, CD320, DMAP1, UBN1, DCP1A, MED4, RIC8A, LUC7L, TIGAR, RTN4, RPR1B, EXOS3, PRD10, ANLN, PDLI7, DDX21, SAR1A, SIAS, SHLB2, MBNL1, AAAS, BAZ1A, ADPPT, OSTC, UBQL4, RAB6B, ABC3C, PHP14, HELLS, KRT84, KRT82, SYFB, PLPL3, BMP2K, ATG3, SMC4, OLA1, ZF64B, RBM12, MDN1, STAU2, T106B, SPS2L, CYRIB, DD19A, MRGBP, CC50A, SEP11, MED17, INT13, DDX18, NLE1, RBM22, WDR70, SLTM, LYAR, MED29, NSMA3, THUM1, TRM1, CARF, GAR1, FA53C, BCLF1, TAB2, FKB11, TMOD3, UGGG1, CDK12, GGA3, TLS1, IF2B1, HPBP1, MAT2B, MYOF, ITSN2, BICRA, EHD2, CNOT2, SPN90, LMCD1, CHMP5, COMD9, CWC15, THYN1, HACD3, KCMF1, RAI14, VAPA, H2AW, TMEM9, SEP10, CACO1, DAPLE, RCC2, MBD5, SLAI2, BCCIP, RRBP1, CPSF2, SYLC, MYEF2, RBM27, KANL3, ANFY1, RERE, ATX10, EP15R, SAE1, COPG2, MRC2, ASH2L, DKK3, TRMB, GULP1, VPS29, GRHPR, CATZ, SAE2, NXF1, PEF1, DNJB4, STK39, CNOT8, ABCF2, GTR6, Z3H7B, TCF20, HMGX4, SWP70, SUN2, RLP24, LIMA1, SRP68, CHRD1, SEPT9, UBQL2, CEIP2, S30BP, MED13, PFD2, PUF60, NRBP, TR112, XPO7, LCAP, MBD1, SIX4, CNOT7, PLXA1, MAGL2, TRM6, FOXD3, DBNL, DDX41, VPS28, CCNL1, TASOR, JUPI1, GMEB2, MRT4, CPSF3, GPTC8, NXT1, RCOR1, RALY, TF3C4, ACINU, AGO2, NUP50, ZHX1, CDV3, CNO11, ZN236, PSME2, MCTS1, ASAP1, MRTFB, ZMIZ1, MED23, PRR12, YETS2, HECD1, PKCB1, COR1C, TPX2, AKP8L, MAFF, TMCO1, EPDR1, DD19B, PPT2, PRP19, SYNP2, UBQL1, VPS4A, G3BP2, CHIP, PACN2, MAGD2, SNX6, SSRG, FAF1, EPCR, CSN3, NSF1C, SCAF8, TRI33, TNR6B, ZC3H4, SMAG1, TRAK1, FOXJ3, SRRM2, PA2G4, SPG7, SMC3, SCML2, ZN148, RTRAF, RUVB2, CDYL, EIF3L, PLAP, RUVB1, NUDC, COF2, SYFA, DRG1, NCKP1, OFUT2, PKHA6, WDR37, E41L3, MYH15, R3HD2, KDM2A, RRP44, RCL1, DDX52, TMA7, YBOX2, MAN1, CHSP1, TR150, WBP11, MED16, NOP58, GIT1, ZN281, SGT1, AAR2, NOSIP, MEMO1, LSM2, SHLB1, LC7L2, SBDS, TMED7, SF3B6, ORN, RRP15, NOP16, PPIL1, MED31, KAD6, STRAP, RTCB, FBX7, NOC2L, RL36, CHTOP, SAMH1, PKP3, SALL2, TLN1, TRRAP, ARIP4, MTCL1, MYO5A, CRBG1, HYOU1, ATG4B, TBL2, KLF12, LSM4, RN114, PRC2C, PPME1, RBM7, YTHD2, PAXB1, SP16H, CD2AP, TNPO3, RBM8A, SNX9, NUBP2, HCFC2, TRUA, SERC, NCOR2, SPIN1, COPG1, GMEB1, CLIC4, CFA20, TACC3, MD1L1, BZW2, EPN1, TEST, IF2B2, S4A7, ROBO1, NCOA3, SCC4, ZHX2, S23IP
Boulard M, Rucli S, Edwards JR, Bestor TH. Methylation-directed glycosylation of chromatin factors represses retrotransposon promoters. Proceedings of the National Academy of Sciences of the United States of America 2020 117(25) 32522876
Abstract:
The mechanisms by which methylated mammalian promoters are transcriptionally silenced even in the presence of all of the factors required for their expression have long been a major unresolved issue in the field of epigenetics. Repression requires the assembly of a methylation-dependent silencing complex that contains the TRIM28 protein (also known as KAP1 and TIF1β), a scaffolding protein without intrinsic repressive or DNA-binding properties. The identity of the key effector within this complex that represses transcription is unknown. We developed a methylation-sensitized interaction screen which revealed that TRIM28 was complexed with O-linked β-N-acetylglucosamine transferase (OGT) only in cells that had normal genomic methylation patterns. OGT is the only glycosyltransferase that modifies cytoplasmic and nuclear protein by transfer of N-acetylglucosamine (O-GlcNAc) to serine and threonine hydroxyls. Whole-genome analysis showed that O-glycosylated proteins and TRIM28 were specifically bound to promoters of active retrotransposons and to imprinting control regions, the two major regulatory sequences controlled by DNA methylation. Furthermore, genome-wide loss of DNA methylation caused a loss of O-GlcNAc from multiple transcriptional repressor proteins associated with TRIM28. A newly developed Cas9-based editing method for targeted removal of O-GlcNAc was directed against retrotransposon promoters. Local chromatin de-GlcNAcylation specifically reactivated the expression of the targeted retrotransposon family without loss of DNA methylation. These data revealed that O-linked glycosylation of chromatin factors is essential for the transcriptional repression of methylated retrotransposons.
Species: Homo sapiens
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Liu Y, Chen Q, Zhang N, Zhang K, Dou T, Cao Y, Liu Y, Li K, Hao X, Xie X, Li W, Ren Y, Zhang J. Proteomic profiling and genome-wide mapping of O-GlcNAc chromatin-associated proteins reveal an O-GlcNAc-regulated genotoxic stress response. Nature communications 2020 11(1) 33214551
Abstract:
O-GlcNAc modification plays critical roles in regulating the stress response program and cellular homeostasis. However, systematic and multi-omics studies on the O-GlcNAc regulated mechanism have been limited. Here, comprehensive data are obtained by a chemical reporter-based method to survey O-GlcNAc function in human breast cancer cells stimulated with the genotoxic agent adriamycin. We identify 875 genotoxic stress-induced O-GlcNAc chromatin-associated proteins (OCPs), including 88 O-GlcNAc chromatin-associated transcription factors and cofactors (OCTFs), subsequently map their genomic loci, and construct a comprehensive transcriptional reprogramming network. Notably, genotoxicity-induced O-GlcNAc enhances the genome-wide interactions of OCPs with chromatin. The dynamic binding switch of hundreds of OCPs from enhancers to promoters is identified as a crucial feature in the specific transcriptional activation of genes involved in the adaptation of cancer cells to genotoxic stress. The OCTF nuclear factor erythroid 2-related factor-1 (NRF1) is found to be a key response regulator in O-GlcNAc-modulated cellular homeostasis. These results provide a valuable clue suggesting that OCPs act as stress sensors by regulating the expression of various genes to protect cancer cells from genotoxic stress.
O-GlcNAc proteins:
RBM47, SBNO1, CNOT1, RGPD3, P121C, PDLI1, MYO1C, AIP, PSD11, PGRC1, TAF4, CLIC1, IPO5, IF2B3, AGRIN, PLOD2, HMGN4, IMA4, PESC, NOP56, DDX3X, PODXL, IMA3, NFIB, ARI1A, G3PT, PDCD5, TCRG1, PSA7, SCAM3, HGS, MYPT1, HNRDL, XPO1, ZN609, SC16A, SR140, SET1A, NPC1, TBX3, ARC1B, TIF1A, PGRC2, PFD6, NKRF, ZN185, OGT1, HMGB3, PPM1G, EIF3D, IPO8, RPA34, NUP42, DHX15, PRP4, SERA, PSMD3, RFOX2, PAPS1, MCA3, HNRPR, PRPF3, TPD54, IF4G3, KLF4, E41L2, DENR, XPOT, PRC1, ZN207, GET3, BUB3, ACTN4, BUD23, SYNC, KRT86, CPSF5, U3IP2, CALU, SAHH2, MED14, SMCA5, ZN862, GANP, KDM1A, ACSL4, SNX3, OGA, HNRPQ, PLOD3, MAFK, IMA7, UGDH, PQBP1, DKC1, IF2P, EDF1, DNJA2, BRD4, PFD1, WDR1, CPNE3, ZC11A, CLU, T22D2, PP6R2, CREST, ANR17, PDCD6, TBCA, H2AY, FLNB, NCOR1, SC22B, PR40A, PSIP1, SRS10, SF3B1, CSDE1, NPM3, U520, NU155, WDHD1, CRTAP, IDHC, CCNK, PIAS2, SPF27, DNJC8, RL1D1, SRP72, MTA2, TOM70, TOX4, SC24D, SUN1, NFAT5, AP2A2, SC31A, SEM3D, AGFG2, ZRAB2, LC7L3, LYPD3, FKBP9, SMC2, IPO7, AHSA1, PSMG1, SC24A, SC24B, CNOT4, OXSR1, HS74L, AP2A1, BAG3, CLPT1, ACL6A, LDHA, AATM, EGFR, PGK1, ASSY, LDLR, K1C14, LMNA, APOA2, FINC, ALBU, TFR1, PROC, ALDOA, CYTB, ANXA1, GCR, KITH, THY1, K2C1, G3P, HSPB1, RPN1, GNAI2, AT1A1, AT1B1, ADT2, IF2A, HMGN2, ICAM1, RLA2, JUN, LA, ITB1, K1C18, K2C8, CDK1, ATPB, S10A6, ENOA, PYGL, G6PI, NPM, TPM3, ITAV, ACBP, LDHB, PDIA1, H10, CATD, ANXA2, TBB5, PROF1, SYEP, HS90A, HNRPC, TSP1, SP1, ANXA6, RHOC, DAF, MDR1, 4F2, HS90B, SRPRA, ASNS, CY1, RU17, ITA5, NFIC, VIME, RS17, K2C7, ANXA5, K1C16, RSSA, SNRPA, GSTP1, LEG1, HMGB1, TPM1, ROA1, RU2A, PARP1, PPBI, UCHL1, ALDOC, ATX1L, HS71B, CALM3, RO60, H14, PTPRF, THIO, ESTD, CH60, BIP, HSP7C, LAMP1, TOP1, TOP2A, PYC, C1TC, MPRI, ADHX, PABP1, PCNA, HARS1, IMDH2, TPR, KCRB, ACTN1, XRCC6, XRCC5, RINI, EF2, K1C10, K2C5, PDIA4, P4HA1, PLST, T2FB, CD59, MIF, GLU2B, CBPM, AK1A1, KPYM, ENPL, CCNB1, PO2F1, HNRPL, SYDC, PLAK, ALDR, AMPN, ERF3A, EZRI, FOSL1, FOSL2, MCP, NQO1, GNS, ZEP1, RS2, DESP, MUC1, CD44, CBR1, CREB1, H15, H13, H12, NCPR, AT2A2, CD36, STMN1, HSP76, HMGA1, JUNB, UBF1, JUND, ATF7, CEBPB, PYRG1, DDX5, PFKAL, LEG3, TCPA, PTN1, RL35A, RL7, VINC, SON, RL17, PGAM1, RCC1, ATF1, ML12A, NUCL, SPEE, RXRA, NFKB1, IF2B, ANXA7, BTF3, PSB1, MPRD, LMNB1, CSRP1, FLNA, 5NTD, VDAC1, CD9, TGM2, PIMT, FBRL, PUR2, PUR6, UBA1, NDKB, ROA2, RFX1, CBL, TCEA1, ITA6, SFPQ, PPIB, SYWC, RS3, NFYA, SAHH, COF1, IF4B, KTHY, EF1B, PPAC, CDK2, MCM3, RS12, BRD2, DNJB1, ATPA, PSA1, PSA3, PSA4, S100P, ITA3, MOES, DDX6, DNMT1, PAX6, U2AF2, RL13, S10A4, HMGB2, PTBP1, SYTC, SYVC, EF1G, STOM, 1433T, ARNT, RL10, RFA1, APEX1, PYR1, CALR, MAP4, CALX, TEAD1, GRN, EPHA2, 3MG, TKT, RBMS1, PML, EF1D, ERP