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Qin H, Chen Y, Tang F, Yue X, Nie Y, Huang W, Ye M. Endo-M Mediated Chemoenzymatic Approach Enables Reversible Glycopeptide Labeling for O-GlcNAcylation Analysis. Angewandte Chemie (International ed. in English) 2022 35289036
Abstract:
To selectively enrich O-linked β-N-acetylglucosamine (O-GlcNAc) peptides in their original form from complex samples, we report the first reversible chemoenzymatic labeling approach for proteomic analysis. In this strategy, the O-GlcNAc moieties are ligated with long N-glycans using an Endo-M mutant, which enables the enrichment of the labeled glycopeptides by hydrophilic interaction liquid chromatography (HILIC). The attached glycans on the enriched glycopeptides are removed by wild-type Endo-M/S to restore the O-GlcNAc moiety. Compared with the classic chemoenzymatic labeling, this approach enables the tag-free identification, and eliminates the interference of bulky tags in glycopeptide detection. This approach presents a unique avenue for the proteome-wide analysis of protein O-GlcNAcylation to promote its mechanism research.
O-GlcNAc proteins:
TM271, HEAT9, SBNO1, P121B, GGT3, TAF4, AGRIN, MEIS1, RNT2, NFIB, MA2B1, CDKA1, ABLM1, ADA10, KMT2D, MYPT1, ZN609, SC16A, PLXB2, SET1A, VA0E1, TNC18, TLR3, TET3, PRPF3, IF4G3, ZN207, AKAP8, CALU, EXTL3, PCDH7, TRAK2, GANP, TSN3, MAFK, KERA, SRGP2, ANR17, NCOR1, U520, CRTAP, ATRN, CBPD, RMP, TOX4, SC24D, SRBS2, SUN1, ERLN2, PCF11, SCAF4, VAPB, SMC2, SC24B, CNOT4, EGFR, HPT, GLHA, LMNA, NU3M, VTNC, GCR, A1BG, K2C1, HSPB1, RPN1, AT1B1, K1C18, K2C8, ITAV, GDN, TRY1, SAP, MET, NFIC, VIME, K2C7, SNRPA, UCRIL, ATX1L, NT2NC, LYAG, LAMC1, PPAL, LAMP1, MPRI, TPR, SKIL, T2FB, GLU2B, NID1, PO2F1, MCP, ZEP1, CD36, ATF7, EGR1, SON, ATF1, ICAL, CO5A1, ASPG, SFPQ, ITA3, NKG2A, ARNT, TEAD1, 3MG, TKT, UFO, HXA11, HXC9, ZEP2, HNRH1, ELF1, CD68, RFC1, FBN1, NU214, RL4, SCNNA, SRP14, NUP62, TAGL2, CUX1, IL6RB, PBX2, STAT3, NBL1, LAP2A, LAP2B, LIFR, PCP, ECE1, MUC18, MATR3, SSRB, VDAC2, ATRX, NOTC1, UTRN, RFX5, HSP13, CCN3, AGRE5, NR2C2, CDK8, CENPF, YLPM1, RBM25, NU153, RBP2, TAF6, EMD, PPT1, SMCA4, HCFC1, AGFG1, NUP98, PTTG, ATX1, AT1B3, AF17, DSRAD, LAMB2, CAD13, ITA1, NU107, TGFB2, ACTG, CXAR, PITX1, CNTP1, ABCA4, PHC1, ADA17, KGD4, SARNP, FOXK1, DAB2, HNRPU, SPTB2, FOXK2, MEF2A, SP3, PLOD1, KMT2A, UPAR, IF4G1, NOTC2, SRS11, SUH, MFGM, AHNK, TMM62, GALT1, BST2, SIA4C, ASPH, BPTF, NFIA, FOXC1, ADAM9, NFAC2, MAMD1, NAB1, TBB3, PKD2, LAMB3, KLF5, NFYC, CDK13, SCRB2, LRP8, VEZF1, UBP2L, LAGE3, MDC1, RRP1B, MEF2D, ARI5B, NUMA1, PON2, RCN1, TAF1C, SF01, MED1, JHD2C, ELF2, TAB1, ZFHX3, ZYX, ADRM1, LAMA4, TAF9, FOXD1, LAMA3, RFX7, QSER1, SVEP1, SYTL3, EPC2, LUZP6, CRTC2, YIF1B, BCORL, K2026, PRC2B, ZN362, LRIF1, UBAP2, RBM26, VP13D, RPRD2, RN220, ZN318, TASO2, ECM29, ARID2, SE1L3, BICRL, SCAR3, NIPBL, LIN54, FSTL4, TET2, ZNT6, GOLM2, NFRKB, S39A4, ZC3HE, FIP1, CRTC3, PLGT2, SUSD1, MCAF1, BCOR, IGS10, B3GLT, CD109, FRRS1, SRCAP, NBEL1, UBN2, RAPH1, HAKAI, ASXL2, SPT6H, KDM3B, NRK, NUP54, POGZ, MAVS, PK1L3, PLGT3, EMSY, RAI1, YTHD3, LDB1, LYRIC, OSTM1, PRSR1, DDX42, P66A, FOXP4, CTL2, TEX2, MGAP, ANKH1, SUGP1, NUP93, PLD6, FNBP4, ARFG1, PMGT2, GOLM1, PGLT1, TM87A, CA131, PHC3, SP20H, ARI1B, NUP35, OR6C4, VA0E2, PLBL2, PUM2, SPP2B, DLG5, GPX8, PO210, ZN384, LMO7, MUC16, P66B, BBX, SMG7, NICA, TM131, PHF3, TAF4B, GGH, GSLG1, FUBP2, LPP, NCLN, TTC17, FKB10, TOM6, PF21A, RBM33, I17RA, P121A, PDLI5, CREL1, FUBP3, TXD15, LOXL4, VCIP1, CHAP1, Z512B, ZFR, EP400, HUTU, RBM14, KI20B, PHF12, PANX1, CIC, MED15, ERBIN, JMJD8, MINT, TEAD3, SEM3C, WAC, DIDO1, HNRL1, TM109, NUP58, M4A8, YTHD1, ULBP3, AMRA1, TANC1, TWSG1, S22A4, SP130, APC1, I2BPL, WNT5B, VP33B, EPC1, ADNP, ZN106, FOXP1, PTN23, WNK1, SIA7D, ZHX3, PIEZ2, SG196, TM231, PEAK1, DOCK5, CF155, CNO10, MLXIP, SIAE, PKHA5, RC3H2, TAF9B, ZBT20, NCOA5, TANC2, ZN532, CELR2, APMAP, ABCB9, CHSTB, UBN1, NECT3, PDLI7, RBM12, OCAD1, CARF, ETAA1, HXC10, TAB2, CELR1, CDK12, CNOT2, CRIM1, TMEM9, RCC2, CHD7, RBM27, KANL3, MRC2, SUCO, TCF20, SUN2, UBQL2, LCAP, GGT7, TASOR, GMEB2, ZHX1, DSE, WWC3, MRTFB, ZBT21, PRR12, YETS2, NOTC3, KMT2B, PRP19, MINP1, SCAF8, ZC3H4, SRRM2, SCML2, UST, ICE1, ZN281, DAAM1, IRS2, PRC2C, NCOR2, NPTN, GMEB1, POMT1, S4A7, S23IP
Species: Homo sapiens
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Ramirez DH, Yang B, D'Souza AK, Shen D, Woo CM. Truncation of the TPR domain of OGT alters substrate and glycosite selection. Analytical and bioanalytical chemistry 2021 34725712
Abstract:
O-GlcNAc transferase (OGT) is an essential enzyme that installs O-linked N-acetylglucosamine (O-GlcNAc) to thousands of protein substrates. OGT and its isoforms select from these substrates through the tetratricopeptide repeat (TPR) domain, yet the impact of truncations to the TPR domain on substrate and glycosite selection is unresolved. Here, we report the effects of iterative truncations to the TPR domain of OGT on substrate and glycosite selection with the model protein GFP-JunB and the surrounding O-GlcNAc proteome in U2OS cells. Iterative truncation of the TPR domain of OGT maintains glycosyltransferase activity but alters subcellular localization of OGT in cells. The glycoproteome and glycosites modified by four OGT TPR isoforms were examined on the whole proteome and a single target protein, GFP-JunB. We found the greatest changes in O-GlcNAc on proteins associated with mRNA splicing processes and that the first four TPRs of the canonical nucleocytoplasmic OGT had the broadest substrate scope. Subsequent glycosite analysis revealed that alteration to the last four TPRs corresponded to the greatest shift in the resulting O-GlcNAc consensus sequence. This dataset provides a foundation to analyze how perturbations to the TPR domain and expression of OGT isoforms affect the glycosylation of substrates, which will be critical for future efforts in protein engineering of OGT, the biology of OGT isoforms, and diseases associated with the TPR domain of OGT.
O-GlcNAc proteins:
UBA6, SBNO1, CNOT1, SMHD1, ADAS, DX39A, BACH, MYO1C, PSD11, PSD12, TAF4, CLIC1, EIF3F, IPO5, DNM1L, AGRIN, PLOD2, IMA4, NOP56, DDX3X, PDXK, ANM5, TCRG1, PSA7, SCAM3, HAT1, HGS, MYPT1, HNRDL, XPO1, PUR4, NPC1, SCAM2, ARC1B, ARPC2, ARPC3, RPAC1, GNPAT, ZN185, SURF4, OGT1, PPM1G, EIF3D, EIF3H, P4HA2, MAGB2, NUP42, DHX15, MCES, SERA, PSMD3, PAPS1, ZW10, HNRPR, TXNL1, E41L2, XPOT, TIM44, PLRG1, ZN207, GET3, BUB3, ACTN4, AP1G1, SNG1, SNG2, SYNC, MYO1B, NRDC, CALU, EDIL3, SAHH2, DHX16, SMCA5, KDM1A, OGA, HNRPQ, PLOD3, DIAP1, TSN3, PLIN3, CTND1, MAGC1, USO1, DKC1, IF2P, DNJA2, WDR1, CPNE3, T22D2, ANR17, H2AY, FLNB, CISY, SC22B, PR40A, ERLN1, SF3B1, CSDE1, U520, NU155, EIF3G, FACE1, IDHC, ATRN, STAM2, PRAF3, FLOT1, RL1D1, WFS1, MTA2, TOX4, SC24D, SUN1, FRYL, NFAT5, GLSK, SC31A, UBR5, LC7L3, FKBP9, SMC2, IPO7, AHSA1, SGPL1, SC24B, ACSL3, CDS2, AP2A1, BAG2, AIFM1, FADS2, CLPT1, TOM40, ACL6A, LDHA, COX2, PNPH, AATM, PGK1, ASSY, FOS, LMNA, FINC, TFR1, NU4M, NU5M, GLCM, FUCO, ALDOA, ANXA1, OAT, TBB4A, G3P, CPNS1, HSPB1, TYSY, RPN1, RPN2, GNAI2, AT1A1, AT1B1, ALDH2, ADT2, PCCA, IF2A, RLA2, RLA0, LA, ITB1, ATPB, IF4E, ENOA, G6PI, NPM, ITAV, HYEP, LDHB, PDIA1, H10, CATD, ANXA2, CAN1, TBB5, TRY1, SAP, PROF1, SYEP, CATB, HS90A, HNRPC, LAMB1, YES, FUMH, TSP1, ANXA6, MDR1, 4F2, PFKAM, HS90B, SRPRA, ASNS, ODPA, RU17, GNAI3, ANXA5, RSSA, CD63, SNRPA, GSTP1, LEG1, CN37, DLDH, ROA1, PARP1, LKHA4, ALDOC, GPHRB, HS71B, LYAG, RRAS, H14, ODP2, THIO, ESTD, CH60, CLUS, BIP, LAMC1, PPAL, HSP7C, GTR1, GTR3, ODPB, PYGB, LAMP1, TOP1, TOP2A, G6PD, PYC, C1TC, MPRI, ADHX, PABP1, PCNA, HARS1, IMDH2, TPR, KCRB, ACTN1, PEPD, XRCC6, XRCC5, LAMP2, RINI, EF2, PDIA4, P4HA1, TCTP, ACPH, ETFA, GLU2B, FPPS, KPYM, ENPL, IDE, HNRPL, SYDC, ALDR, B4GT1, EZRI, FOSL2, NDKA, GNS, RS2, RFA2, BGAL, H12, AT2A2, FAAA, AATC, JUNB, ITA2, CAN2, PYRG1, DDX5, PFKAL, PRS6A, TCPA, RL35A, ARF4, RL7, VINC, SON, RL17, PGAM1, DNLI1, PLCG1, NUCL, HXK1, RPB3, E2AK2, TFE3, SPEE, IF2B, ANXA7, PSB1, MPRD, LMNB1, VATB2, FLNA, VDAC1, TGM2, FBRL, PUR2, PUR6, UBA1, NDKB, ADRO, ROA2, RFX1, QCR2, SFPQ, PPIB, SYWC, RS3, SAHH, COF1, IF4B, PPAC, RPB1, MCM3, RS12, DNJB1, ATPA, PSA1, PSA2, PSA3, PSA4, ITA3, MOES, DDX6, DNMT1, U2AF2, RL13, PTBP1, SYTC, SYVC, EF1G, STOM, RL10, RFA1, APEX1, CD82, PYR1, CALR, MAP4, CALX, PSB8, PSA5, PSB4, PSB6, PSB5, DPOD1, TEAD1, MK01, AMPL, IMPA1, EPHA2, TKT, EF1D, PRDX6, BLVRB, PRDX5, PRDX3, RL12, ECHM, PEBP1, PDIA3, 2AAA, CDC27, PURA2, AMRP, PUR8, CLIP1, SL7A1, RPB2, SDHA, GDIA, METK2, DNJA1, QCR1, PUR9, HNRH3, HNRH1, 1433B, 1433S, STIP1, PRDX2, RL9, KINH, MRP1, MCM4, MCM5, MCM7, GLYM, HSP74, CTNA1, CTNB1, PHB1, RADI, RFC4, RL22, FBN2, GDE, MYH9, MYH10, COPB2, SOAT1, ADDA, BASI, FUS, NU214, PRS7, MP2K2, ATPG, HEM6, RL4, PGM1, ODO2, SRP14, NUP62, FDFT, TAGL2, TALDO, RBMX, VKGC, VATA, GRP75, IF4A3, RS19, RL3, OST48, AN32A, CAP2, TCPZ, RL13A, STAT3, MDHC, MDHM, ECHA, IF2G, KDM5C, GARS, SYIC, STAT1, CASP3, LPPRC, SATT, LIS1, MUC18, MATR3, MSH2, MAGA4, RANG, NAMPT, PRS6B, PPIC, VDAC2, UBP5, KI67, RAGP1, RECQ1, NOP2, CRKL, BAG6, NSF, RL27A, RL5, RL21, RL28, RS9, RS5, MAP1B, UTRN, IQGA1, STT3A, CAPZB, SYQ, UCRI, PPCE, COPD, LSS, CD151, PSMD8, TCPE, NEST, HSP13, PIPNB, PAXI, AL9A1, RL34, LMAN1, NASP, FAS, TCPG, EFTU, AL7A1, SYAC, SYCC, SYSC, PRI2, PSB3, PSB2, MCM2, ACADV, YLPM1, TMEDA, RBM25, NU153, RBP2, NDUV1, GSK3A, TAF6, GUAA, GDIB, EMD, SERPH, VASP, TCPQ, TCPD, ANX11, PAPOA, FXR1, RAB5C, RAB7A, RT29, SMCA4, BAP31, HCFC1, DHB4, HDGF, ROA3, 6PGD, HNRPM, IMA1, GDIR1, AGFG1, HNRPF, MSH6, RBM5, SPSY, CAZA1, NUP98, BIEA, PPP5, ACLY, MAP11, COPB, COPA, SC5A3, SMTN, MOT1, IST1, SC24C, SYRC, SYYC, UBP14, BCAT1, AT1B3, RD23B, P5CS, IF5, S12A2, XPO2, TERA, ECHB, AFAD, NP1L1, DSRAD, SEC13, PSA, HNRH2, EIF3B, ATPK, SYMC, IF6, CTBP2, TMM33, GEMI4, CORO7, NU107, ARPC4, TPIS, EIF3E, PP4C, IF4A1, PSA6, CDC42, DEST, GMFB, SRP54, RAB10, UBC12, ARP3, ARP2, ABCE1, RAP2B, RS3A, RL15, RL37A, S61A1, DAD1, NPC2, HNRPK, 1433G, RS7, PRS4, PRS8, RS8, RS15A, RS16, 1433E, RS14, RS23, RS13, RS11, SMD2, SMD3, PRS10, RL7A, ERF1, CNBP, RS4X, PP2AB, RS6, H4, RAB1A, RAN, RL23, RS25, RS28, RL30, RL31, RL10A, RL32, RL11, RL8, PPIA, TRA2B, AP2B1, 1433Z, IF5A1, RACK1, YBOX1, UB2L3, TBA4A, IF4G2, GTF2I, TCPB, PRKDC, RL24, SRSF3, FOXK1, DAB2, RBM10, MPCP, VIGLN, CLH1, FKBP3, HNRPU, U2AF1, SPTB2, AMPD2, CAP1, HMCS1, LAT1, EXOSX, PFKAP, EWS, PLCB3, ODO1, RL18A, MP2K1, FKBP4, PLOD1, RL6, TOP2B, DYST, KMT2A, LMNB2, CEBPZ, TF65, IF4G1, LGUL, 1433F, DYN1, EF1A2, PTN11, PUR1, GFPT1, EXOS9, SUH, GABPA, PRDX1, RL18, C1QBP, CKAP4, KHDR1, LRP1, SRSF1, PP2BA, DHX9, GOGA2, LG3BP, CD47, SSRP1, NSUN2, NCBP1, EP300, AHNK, MGAT2, GALT2, AP1B1, MPPA, SCRN1, NU160, SCAP, TBL3, TF3C1, ASPH, TROP, BPTF, AIMP1, ILF2, ILF3, LMAN2, TRAP1, FLII, ACACA, CSN1, MTAP, EI2BE, TADBP, ROA0, PRDX4, PSMD2, SRSF6, TIF1B, DHRS2, G3BP1, MTA1, EIF3I, UB2V1, DC1I2, NNTM, TCOF, TMED1, VPP3, PICAL, RIPK1, CUL2, CUL4A, CUL4B, FCL, MOGS, CD166, SPTN1, DX39B, BLMH, CKAP5, COTL1, HNRPD, SCRB2, EIF3A, UBP2L, SCRIB, FHL2, DPYL3, DCTN1, DYHC1, SRC8, FLOT2, TRI25, FLNC, GALE, CAPR1, RBM39, MCM6, TRIPC, PUM1, EPN4, RRP5, BMS1, UBP10, GANAB, LBR, MVP, MEF2D, CHD4, LASP1, GPNMB, ZN638, IMB1, NUMA1, CND2, SPCS2, PSMD6, ABRX2, SEPT2, CND1, U5S1, SYK, WDR43, PDIA6, PA1B3, IPYR, TEBP, NONO, PWP2, RCN1, L2GL1, TMED2, PCBP1, PCBP2, SF3B3, SC23A, SF3A1, TSN, SF01, PCH2, MED1, MARE1, ELAV1, NSDHL, TAB1, AAAT, MLF2, VAS1, ZYX, SEPT7, ADRM1, CCDC6, UAP1, PSMD5, PKN2, DDB1, DPYL2, KCC4, SRSF7, DREB, NRF1, FSCN1, IF16, PCKGM, UPP1, UGPA, TRXR1, HNRL2, INF2, PDS5A, QSER1, QRIC1, SMU1, P3H1, K2C71, TIM50, GXLT1, TB10B, SNUT2, HERC4, P4R3B, EOGT, CE170, UBR4, SKT, UBAP2, RBM26, RPRD2, ZMYM4, TWF2, LIN54, RHBD2, MBOA5, EDC4, PRP8, SCYL2, NFRKB, TTC37, NCEH1, BRAT1, ZC3HE, C1TM, SBSN, CREL2, PACS1, GGYF2, SRCAP, TMTC3, IKIP, SPT6H, SND1, DDX46, BZW1, DHX30, EIF3M, CYFP1, KDM3B, RIR2B, ERMP1, ZCCHV, NUP54, POGZ, MYH14, CLAP1, EMSY, SH3R1, HUWE1, TMED4, SYVN1, ZN598, PHLB1, LUZP1, CAND1, TSYL5, EZHIP, CARM1, DDX42, P66A, C2CD5, I2BP1, RB6I2, PLD3, P3H3, WDR75, Z3H7A, ANKH1, CCAR1, DJC10, SMAP1, SPART, RPTOR, NUP93, ZN687, SYNPO, FNBP4, CPSF7, EMC1, ENAH, S43A3, GT251, SUMF2, PGLT1, TXND5, PAIRB, LS14A, ABCF1, FBX22, NEUA, CADM4, ATLA2, PLBL2, WDR36, SMRC2, NPL4, S35B2, UBA3, TBC15, CIP2A, STT3B, PNPT1, HM13, DTX3L, PO210, GEMI5, PARD3, IPO4, SCRB1, RAB2B, NU133, PDC6I, S20A1, ATX2L, PALLD, DDX1, GBF1, NICA, TM9S4, HS105, GCN1, PRP16, NU205, PXDN, TFG, ARC1A, HDAC2, CBP, SYMPK, DDX17, CELF1, UFD1, GSLG1, RENT1, TNPO1, GLMN, USP9X, UBP7, SCAM4, SMCE1, MYDGF, PIGT, NCLN, EXOC4, FERM2, FUBP1, FKB10, APH1A, PPWD1, AP2M1, RCN3, ELP4, SEH1, DNJA3, CYFP2, PGM2, PDLI5, ERO1A, FUBP3, ITCH, TMX3, CNDP2, ZFR, EP400, SNX27, PRRC1, ZN512, MBOA7, DOCK7, ARAP1, IPO9, RBM14, NED4L, VPS35, CIC, MED15, EFGM, MCCA, MYADM, SIN3A, MINT, PNKP, NIBA2, TBCB, PSB7, CNN2, PSMD1, EYA3, SORT, NUP88, P2RX4, DNJC7, PHB2, KIF2C, ATX2, HCD2, ROAA, NP1L4, ACON, TM9S2, S29A1, TS101, TCPH, ANM1, CND3, MBB1A, COR1B, GCP2, ESYT1, DIDO1, MCMBP, TBCD, TBB6, RPC3, HNRL1, HTAI2, TMED9, NUP58, SFXN3, OSB10, NAA15, S12A9, SRRT, EIF2A, WDR11, FTO, TTYH3, WDR12, YIPF3, PITH1, PAIP1, NAT10, XRN2, TOLIP, SP130, BRD8, DDX47, MAGT1, RAB1B, LMA2L, I2BPL, GHITM, TMX1, PTN23, OFUT1, PIGU, WNK1, AMPB, HEAT1, NOL6, WDR26, AT133, ACAD9, GORS2, MARH7, SFXN1, PKHA5, XPO5, MYG1, EMAL4, TM9S3, APMAP, ARFG3, RTN4, ANLN, XPP1, PDLI7, DDX21, EI2BG, MBNL1, HEBP1, SERC1, F1142, SYFB, SYIM, SMC4, OLA1, RBM12, T106B, CYRIB, DCA13, CC50A, SEP11, FANCI, ARL8B, DDX18, TM160, MIO, CARF, DPP3, RM39, UGGG1, ERAP1, HPBP1, MAT2B, MYOF, ITSN2, HACD3, SEP10, RCC2, DIP2B, CT2NL, RRBP1, SYLC, RBM27, SUCB1, MRC2, DHCR7, DJB11, SAE2, NXF1, SUN2, SRP68, SEPT9, UBQL2, PCYOX, CEIP2, S30BP, MED13, PUF60, NRBP, VATH, IPO11, XPO7, BAZ2A, GGT7, NAGK, DBNL, CDC23, STML2, PARP4, NUDT5, TF3C4, ACINU, RAB21, DNPEP, ASAP1, MRTFB, MED23, HECD1, PKCB1, COR1C, MYO6, PRP19, MAGD2, PSD13, MINP1, NOVA2, WDR3, S12A4, MACF1, SCAF8, TRI33, SMAG1, XCT, SRRM2, PA2G4, SMC3, RUVB2, EIF3L, PLAP, RUVB1, VDAC3, ERGI3, SC5A6, DRG1, NCKP1, AT11B, E41L3, CHSP1, NOP58, ZN281, SYYM, TMX2, SHLB1, SBDS, TMED7, STRAP, RTCB, FBX7, RBGP1, NOC2L, TLN1, DCAF1, HYOU1, PRC2C, YTHD2, SP16H, TNPO3, MAGD1, SNX8, TRUA, NPTN, S12A7, COPG1, BIG1, BZW2, DC1L1, IF2B2, ZNT1, S4A7, ZHX2, S23IP
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, ERP29, PRDX6, RL12, KCY, PEBP1, PDIA3, 2AAA, NMT1, PURA2, UFO, SORCN, ILEU, RPB2, METK2, TIA1, ZEP2, DNJA1, PUR9, HNRH3, HNRH1, 1433B, 1433S, STIP1, S10AB, L1CAM, PRDX2, CDD, ELF1, RL9, CD70, KINH, CSTF2, MCM4, MCM5, MCM7, GLYM, HSP74, PROF2, PHB1, SPB6, RFC4, RL22, K1C9, MYH9, MYH10, COPB2, BASI, FUS, NU214, DEK, K22E, PRS7, ATPG, RL4, PP1G, GNL1, SRP14, NUP62, TAGL2, TALDO, RBMX, VKGC, GRP75, IF4A3, RS19, RL3, OST48, FEN1, CAPG, TXLNA, TCPZ, RL13A, STAT3, MDHC, MDHM, IF2G, GARS, SYIC, LAP2A, LAP2B, STAT1, MTREX, RS27, LPPRC, RL35, CDN2A, ECE1, LIS1, MUC18, MATR3, MSH2, SSRA, RANG, VDAC2, CBX5, UBP5, KI67, RAGP1, RECQ1, NOP2, BAG6, NOTC1, RL27A, RL5, RL21, RL28, RS9, RS5, RS10, IQGA1, CAPZB, IF1AX, RL29, SOX9, COPD, GSH0, PSMD8, PRC2A, TCPE, PTSS1, K2C6C, AGRE5, PAXI, RL34, LMAN1, NASP, FAS, CDK8, TCPG, EFTU, SYAC, SYSC, MCM2, ACADV, YLPM1, TMEDA, RBM25, HINT1, NU153, RBP2, TAF6, GUAA, CRIP1, GDIB, EMD, SERPH, F10A1, MAP2, RL14, TCPQ, TCPD, ANX11, PAPOA, FXR1, FXR2, RAB7A, SMCA4, SSRD, HCFC1, HDGF, ROA3, 6PGD, HNRPM, IMA1, GDIR1, AGFG1, HNRPF, MSH6, CAZA1, CRIP2, NUP98, ACLY, COPA, SC24C, TCP4, SYRC, ATX1, ATN1, SYYC, UBP14, AT1B3, RD23B, SNAA, IF5, PSMD4, XPO2, TERA, AF10, AF17, NP1L1, ADK, DSRAD, SEC13, NH2L1, PSA, EIF3B, SYMC, IF6, CTBP2, TMM33, NU107, EPIPL, TPIS, EIF3E, SC61B, MYL6, ACTB, IF4A1, RS20, PRPS1, PSA6, S10AA, CDC42, DEST, RAB10, UBC12, UBE2N, ARP3, ABCE1, RS3A, RL26, PSME3, RL15, RL27, RL37A, S61A1, PFD3, B2MG, DAD1, SUMO2, WDR5, NTF2, HNRPK, 1433G, RS7, PP1A, PP1B, RS8, RS15A, RS16, 1433E, RS14, RS23, RS18, RS29, RS13, RS11, RUXE, SMD1, SMD2, SMD3, PRS10, RL7A, ERF1, CNBP, RS4X, RL23A, RS6, H4, RAN, RL23, RAP1A,