JPOST Repositoryapplication/xmlhttps://storage.jpostdb.org/JPST001923/files/140605_hata_140605_oe_02_refseq_human8_P_CarbamidoM_PRS.xlsxhttps://storage.jpostdb.org/JPST001923/files/140605_oe_02.rawprimary200OKProteomicsMasaaki OyamaHomo Sapiens (human)https://repository.jpostdb.org/entry/JPST001923The Institute of Medical Science, The University of TokyojPOSTGrowth factor-induced, ERK-mediated induction of immediate-early genes (IEGs) is crucial for cell growth and tumorigenesis. Although IEG expression is mainly regulated at the level of transcription elongation by RNA polymerase-II (Pol-II) promoter-proximal pausing and its release, the role of ERK in this process remains unknown. Here, we identified negative elongation factor (NELF)-A as an ERK substrate. Upon growth factor stimulation, ERK phosphorylates NELF-A, which dissociates NELF from paused Pol-II at the promoter-proximal regions of IEGs, allowing Pol-II to resume elongation and produce full-length transcripts. Furthermore, we found that in cancer cells, PP2A efficiently dephosphorylates NELF-A, thereby preventing aberrant IEG expression induced by ERK-activating oncogenes. However, when PP2A inhibitor proteins are overexpressed, as is frequently observed in cancers, decreased PP2A activity combined with oncogene-mediated ERK activation conspire to induce NELF-A phosphorylation and IEG upregulation, resulting in tumor progression. Our data delineate previously unexplored roles of ERK and PP2A inhibitor proteins in carcinogenesis.ERK-mediated NELF-A phosphorylation promotes transcription elongation of immediate-early genes by releasing promoter-proximal pausing of RNA polymerase II.Ohe Seina S, Kubota Yuji Y, Yamaguchi Kiyoshi K, Takagi Yusuke Y, Nashimoto Junichiro J, Kozuka-Hata Hiroko H, Oyama Masaaki M, Furukawa Yoichi Y, Takekawa Mutsuhiro MPhosphorylations, Wolf-Hirschhorn syndrome candidate 2 protein, phosphorylation., mWHSC2, WHSC2, NELF-A, Wolf-Hirschhorn syndrome candidate 2 homologRibonucleic, PP2A[[C]], DmErk, extracellular signal-regulated kinase activity, PR53, pp44mapk, Materials, PR55, Tyro5, l(3)S026326, RNA pol II, PP2A[B], pr53, anon-WO0118547.420, growth and development, Tumor, RpII215, phosphorylation, DmelCG7109, B', TH1 protein, Anon-66Da protein, LeMPK3, l(3)S031807, RNApol2, PP2A 28D, p44mpk, COBRA1 protein, Dp38, PP2A[C], 5, SAPK2, Fs(3)Hor, l(3)01436, SEM, Sem, Non Polyadenylated, l(3)S110815, MAP-k, RNApolII, Pp2A, pp42, PP2a, l(3)S026226, EK5, l(3)S027313, l(3)S141309, aar, Dsor2, metabolic process resulting in cell growth, NTef2, sem, pp2a, l(3)02414, DmelCG6235, DmelCG9984, PR65, l(3)S035505b, Transforming Genes, decreased, PP2A, Role Concepts, l(3)S048006, cellular growth, CG1554, Pp2A-28D, l(3)S048013, transcription from bacterial-type RNA polymerase promoter, Pol II0[ser2], PP2A[A], Tumors, ERK-A, l(3)S067915, dpERK, dpErk, anatomical protrusion, metabolism resulting in cell growth, negative elongation factor complex, CG7901, ERK-2, DmMAPK, PolII, PMK-2, l(3)S091905, dp-ERK, PMK-1, PP2-AB, PMK-3, Fs(3)Sz11, serine/threonine protein phosphatase 2A, dNelf-E, Role Concept, Benign, 1466/06, Role, pMAPK, pMapK, NELF-D protein, DmPp2A-28D, l(3)S029403, Ribonucleic acids, l(3)S033903, activation, CG7913, Acid, Pol-IIa, DmERKA, rl/MAPK, Wrd, l(3)S042630, PolIIo, RDBP protein, RNAP II LS, Benign Neoplasms, pp2A-B', l(2)41Ac, human, Pol II0[ser5], C78273, Malignant Neoplasms, CT34260, dPP2A A, l(3)S075902b, Oncogene, dpERk, wrd, A/PR65, antagonists and inhibitors, Material, l(3)S066813, H5, l(3)S027127, SAPK, CG2684, B56-2, dRPB1, dRpb1, l(3)S042629, other neoplasm, DmelCG3284, B56-1, 9030612K14Rik, dRPB9, TH1-like (Drosophila) protein, Jacob, l(3)S063110, atypia, LD02456, dPP2A-B56-1, Neoplasms, dPP2A-B56-2, 12559, CG5643, EK2-1, II, l(3)S132907, Pol II, AA407128, l(3)S069206a, l(3)S023141a, Gene Products, Prkm1, CG17291, atypical, i234, l(3)S119908, ERK, Erk, PRKM1, l(3)j11C8, PRKM2, Wolf-Hirschhorn syndrome candidate 2 protein, l(3)S046918, Transforming Gene, PP2A[B'-1], l(3)S088513, Pol II CTD, dPP2A-Balpha, PCBC, NELF, RNAPII0, dPP2A-C, 5559, BEST:LD02456, erk, Neoplasias, PP2A[B'-2], DPR65, L5, l(3)S029110, l(1)DF912, rll, Pol IIo[ser5], CG5994, STK26, p38-2, PP2Ac, Radin blood group protein, ppp2r2c, myelin basic protein kinase activity, Cancer, l(3)S076415, inhibitors, Pol IIo, l(3)S023938, RNA, DPR55, CG 7913, DRT, Malignant Neoplasm, EP3559, Pol IIa, Proteins, stress-activated kinase activity, Phosphorylations, MAP-2 kinase activity, RNS, cg5994, defective, RNAPII, Cell, Concept, MT, DmelCG5643, CG3284, stress-activated protein kinase activity, mapk2, mapk1, Neoplasm, PP2A C, PP2A A, Nelf, INSDC_feature:regulatory, DmelCG5994, l(2)02496, primary cancer, CG12559, mWHSC2, DmelCG17291, P42MAPK, dpERK1, Pol IIo[ser2], postnatal growth, negative elongation factor E protein, Carcinogeneses, l(2)s5286, l(3)S032303, l(3)S075110, Ribonucleic Acid, Cancers, Lds, malignant tumor, tws/aar, l(1)10Ca, l(3)S105605, Gene Proteins, Pol II[ser2], AU018647, dpMAPK, ppp2r2b-a, Jac, CTD, Nelf-e, RNA PolI 215, General activity, SAP kinase activity, NELF-C protein, Neoplasia, Pp2A29B, l(3)S025913, Activity, l(3)Z23, DmelCG7913, Mpk2, postnatal development, 8WG16, Pol-II, p42mapk, trihydrophobin 1 protein, l(3)S060804, SR2-1, l(3)S024838, l(3)S025806, Tumorigenesis., RNA polII, l(3)S045519, l(1)L5, Roles, l(3)S043008b, POL, RNA pol IIo, Concepts, Tws, 0318/07, PP2, l(3)S031006, mitogen-activated protein kinase activity, l(3)S053011, Pol IIo[Ser2], Transforming, l(3)S080409, mpk1, present in fewer numbers in organism, NELF-B protein, negative elongation factor (NELF), RNA Gene Products, DmelCG2684, Erk/Map kinase, l(3)s1801, l(3)S049902, non-developmental growth of a unicellular organism, l(3)S067109b, TYPE 2A SERINE/THREONINE PROTEIN PHOSPHATASE, DERK-A, Oncogeneses, MTS/PP2A, E(sina)7, Rl, MP kinase activity, DERK, v158, malignant neoplasm, R75353, MBP kinase II activity, F20P5.30, polII, NELF-E protein, dERK, Malignancies, ER2-6, negative elongation factor E, l(3)S043029, F20P5_30, negative elongation factor D, l(3)S024834a, tw, negative elongation factor A, PP2a 28D, MAP kinase 2 activity, negative elongation factor C, negative elongation factor B, major histocompatibility complex protein RD, aberrant, Mapk, ribose nucleic acid, l(3)S025832, l(3)S022440, ribonucleic acids, CG7109, Erk1, ERK1, ERK2, mapk1a, l(1)DC912, B/PR55, Erk2, l(3)S101413b, Pp2A-85F, l(3)S023309, TH1L protein, negative elongation factor NELF, p41mapk, l(3)S110008a, mapk1b, MapK, MAPK, non-developmental cell growth, Ribonukleinsaeure, l(3)88Be, Genetic Materials, pentosenucleic acids, erk2, mapk, Genetic Material, BcDNA:LD34343, pol II, l(3)S029701a, ptpa, PP2A B', ERKa, DmelCG1554, BcDNA:RE08694, growth pattern, RNA Pol II, Rpb1, RPB1, l(2R)EMS45-39, non-developmental growth, CAPB, PTPA, DmelCG12559, p38, CG6235, l(3)S061915, CG9984, CG33297, early, DpErk, DpERK, l(3)S066017, Pol II Ser5P, l(3)S032708c, ErkA, ERKA, l(3)S023206, Drosophila, Oncogenesis, spine, Horka, p41, bacterial transcription, p40, l(3)S134601a, Fs(3)Horka, l(3)S022361, Cistron, RPII15, l(3)S061805, pERK, Tumorigeneses, Nelf-E protein, GroupII, Benign Neoplasm, PP2A-29B, Gene, Wdb, dPP2A, l(3)S075515b, l(3)S111515, Malignant, CG13383, protrusion, dPR55, Hek5, l(3)S048507, reduced, subnumerary, DmF2, p42-MAPK, HH9, dB56-2, dB56-1, BcDNA:GM05554, lod, PP2A-A, B'/PR61, PP2A-B, l(3)S022205b, PP2A-C, Genetic, growth of cell, Malignancy, l(3)S146606, RNAP, xp42, RPII215, inhibiteur, decreased number, p42 mitogen-activated protein kinase activity, CG18732, cofactor of BRCA1 protein, ATP:protein phosphotransferase (MAPKK-activated) activity, Ubl, Non-Polyadenylated RNA, RNAP II, inhibidor, negative elongation factor proteins C and D, l(3)S024455, RNA Pol II CTD, DmERK-A, anon-66Da, l(1)G0040, RNA PolII, ert1, l(3)S060203, Mts, l(3)S023013, prkm2, prkm1, CT39192, inhibitor, l(3)S028707, WHSC2, Cistrons, l(3)S052810, development, p38delta, cell expansion, yeast nucleic acid, Protein, MBP kinase I activity, Rpll215, Xp42, antagonists, ribonucleic acid, Genes, mitogen activated kinase activity, Su(Raf)2B, Non Polyadenylated RNA, Non-Polyadenylated, EY2-2, MAPK2, RpII, NELF-E, Protein Gene Products, NELF-D, D-ERK, ERT1, NELF-A, EPHT3, Wolf-Hirschhorn syndrome candidate 2 homolog, 2414, 6330556D22Rik, growth, MAP kinase 1 activityDmErk, extracellular signal-regulated kinase activity, pp44mapk, Materials, GroupII, Tyro5, Mpk2, 12559, p42mapk, Gene, phosphorylation, EK2-1, SR2-1, Hek5, AA407128, LeMPK3, p44mpk, Prkm1, Dp38, mitogen-activated protein kinase activity, DmF2, p42-MAPK, SAPK2, Fs(3)Hor, mpk1, SEM, Sem, ERK, Erk, PRKM1, lod, MAP-k, PRKM2, Wolf-Hirschhorn syndrome candidate 2 protein, DmelCG2684, Erk/Map kinase, Genetic, pp42, EK5, xp42, PCBC, DERK-A, Dsor2, E(sina)7, NTef2, sem, Rl, p42 mitogen-activated protein kinase activity, CG18732, ATP:protein phosphotransferase (MAPKK-activated) activity, erk, MP kinase activity, DERK, DNA Dependent RNA Polymerase II, MBP kinase II activity, rll, DmERK-A, dERK, STK26, p38-2, transcription from bacterial-type RNA polymerase promoter, myelin basic protein kinase activity, ERK-A, ert1, MAP kinase 2 activity, RNA Pol II., dpERK, dpErk, DRT, Mapk, ERK-2, prkm2, stress-activated kinase activity, prkm1, Erk1, CT39192, Phosphorylations, MAP-2 kinase activity, ERK1, ERK2, mapk1a, Erk2, DmMAPK, PMK-2, dp-ERK, PMK-1, WHSC2, Cistrons, PMK-3, Fs(3)Sz11, p38delta, p41mapk, mapk1b, MapK, MAPK, stress-activated protein kinase activity, mapk2, mapk1, MBP kinase I activity, pMAPK, pMapK, Genetic Materials, erk2, mapk, Genetic Material, INSDC_feature:regulatory, Xp42, ERKa, DmERKA, CG12559, rl/MAPK, BcDNA:RE08694, mWHSC2, l(2R)EMS45-39, mitogen activated kinase activity, P42MAPK, Su(Raf)2B, CAPB, dpERK1, DmelCG12559, p38, EY2-2, l(2)41Ac, MAPK2, Lds, early, C78273, DpErk, DpERK, ErkA, ERKA, AU018647, CT34260, D-ERK, dpERk, dpMAPK, ERT1, Material, Horka, p41, NELF-A, bacterial transcription, p40, EPHT3, Wolf-Hirschhorn syndrome candidate 2 homolog, SAPK, CG2684, Fs(3)Horka, RNA Polymerase B, Cistron, DNA-Dependent RNA Polymerase II, SAP kinase activity, pERK, 9030612K14Rik, MAP kinase 1 activityliquid chromatography tandem mass spectroscopy, LC-MS2, 293T Cell, 12-O-tetradecanoylphorbol 13-acetate, 293 Cells, phorbol 12-myristate 13-acetate, determination, PLAT, LC-MS/MS, Human Embryonic Kidney Cell Line 293, Alteplase, Human Kidney Cell Line 293, HEK293 Cell, WHSC2, HEK 293 Cell, Cell, LC-MS-MS, 293T, LC/MS/MS, Tissue-type plasminogen activator chain A, Tissue-type plasminogen activator chain B, tetradecanoylphorbol acetate, T-PA, LC-MSMS, chemical analysis, HEK 293, PATISS, Reteplase, phorbol 12-tetradecanoate 13-acetate, D8Ertd2e, AU020998, Wolf-Hirschhorn syndrome candidate 2 protein, Tpa, TPA, LCMSMS, mWHSC2, reteplase, t-plasminogen activator, HEK293, taurine:pyruvate aminotransferase activity, t-PA, HEK 293 Cell Line, tPA, PMA, 12-tetradecanoylphorbol 13-acetate, HEK293., phorbol-12-myristate-13-acetate, HEK 293 Cells, liquid chromatography-tandem mass spectroscopy, NELF-A, Wolf-Hirschhorn syndrome candidate 2 homolog, alteplase, 3.4.21.68, Cells, HEK, 293T Cells, liquid chromatography tandem mass spectrometry, assay, AW212668, 293 CellfalseMS-based determination of NELF-A phosphorylation LC-MS/MS analysis of Flag-NELF-A purified from TPA-stimulated HEK293 cellsTue Nov 15 00:00:00 GMT 2022PXD038094960636463234