Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/proteinGroups.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/OxidationMSites.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/msmsScans.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/peptides.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/parameters.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/modificationSpecificPeptides.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/msms.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/mzRange.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/msScans.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/MethylKRSites.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/evidence.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/summary.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/allPeptides.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/tables.pdfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/mqpar.xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/20161108_QEX_EM_H4_s8_ArgC.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/20161108_QEX_EM_H4_s6_ArgC.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/20161108_QEX_EM_H4_s7_ArgC.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/03/PXD008965/20161108_QEX_EM_H4_s5_ArgC.rawprimaryOK200imhof@lmu.deAndreas SchmidtMass SpectrometryShotgun proteomicsProstate CancerNot availableLysine methylationChromatinH4k12me1Kmt9Trmt112Methyl-silacMethyltransferaseProstate cancerhttp://www.ebi.ac.uk/pride/archive/projects/PXD008965In vitro methylation assays Samples containing purified Histone H4, purified Methyltransferase and isotopically coded S-adenosylmethionine were digested with ArgC according to the manufacturers instructions. The resulting peptide mixture were desalted using stage tips with C18 material (Empore C18, 3M), dried in a speedvac and redissolved in 0.1% TFA for LC-MS analysis. LC separations were conducted with a linear gradient from 3 to 40% acetonitrile in 50 min and the eluting peptides were directly infused into the mass spectrometer via a nanoESI source. The Orbitrap HF was run in data dependent acquisition mode with one survey scan at 60.000 resolution and up to 10 MS/MS scans at 15.000 resolution. FT spectra were recalibrated using up to three lockmasses for silicones from the ambient air. MS/MS precursor ions required a charge state between 2+ and 6+. For MS/Ms acquisition, the underfill ratio was set to 5%, the normalized collision energy to 27 and the isolation width to 1.5 m/z. Previously seleced precursors were exclude for 20 sec from repeated ananalysis.PrideTICdimethylated residuemonomethylated residueacetylated residueMS raw data were searched for elution profiles of methylated histones peptides with light (12C1H2, +14 Da) or heavy (13C2H2, +18 Da) label and MS/MS spectra were used to localize the modification within the peptide sequence in comparison of the MS/MS data of the unmodified peptide. Proteomic data were searched against a human database to confirm the protein presence and elution time of histone peptides.ProteomicsAxel ImhofQ ExactiveProtein Analysis Unit ZfP LMU Munich BioMedical Center Grosshaderner Strasse 9 82152 Planegg-Martinsried Tel. +49 89 2180 71804PARTIALHomo Sapiens (human)Not availableA.Schmidt@med.uni-muenchen.deBiologicalBiomedicalCECAD, University of Cologne, AG KrügerGermanyIPP2A2, MeCN, Selb., NCMe, ion, determination, developmental stage, precursor, thomson, CG7826, PHAPII, mass-to-charge ratio, 5730420M11Rik, peptide, Histone H2b, Polypeptides, Histone H2a, S Adenosyl L Methionine, ClvPrd, peptido, template-activating factor I, S Amet, CH3-C#N, AdoMet, B1, CG7835, SEC, CG42273, Min, FO-1561, Histone H3.3, DmelCG42273, Ademetionine, T6K12_14, SET, proportion, peptides, FO 1561, TAF-I, iones, phosphatase 2A inhibitor I2PP2A, ipp2a2, sec, min, 2pp2a, mAPC, ions, SCAN, CG10574, F8N16.2, AI047805, DmelCG4299, set, IGAAD, NAGSA dehydrogenase activity, SAM-e, 2PP2A, DmelCG10574, taf-ibeta, D1, Histone H5, dSET, dSet, Histone H4, Histone H7, peptidos, stage, z, Methylations, Histone H1, PTHB1, Histone H3, inner salt, ratio, Gumbaral, (3S)-, phapii, 5'-((3-amino-3-carboxypropyl)methylsulfonio)-5'-deoxy-, DYRK1, data, S-Adenosyl-L-Methionine, 1-(14)C-labeled, 3H-labeled, histone H4, F8N16_2, igaad, Dmel_CG7826, StF-IT-1, Th, LysY, Mischung, acetonitrile, Histone, Peptide, polypeptide, secret agent, Samyr, Silicone, Dm1, Ionen, I-2PP2A, C18, chemical analysis, Dm I-2, I2PP2A, Dyrk1, methylation, AGPR, ACETONITRILE, Dmel_CG7835, cyanomethane, T6K12.14, Mnb, MNB, parent ion, AU020952, HLA-DR-associated protein II, DI-2, FO1561, I-2Dm, CC1, proportionality, ArgC, rate, ethanenitrile, CG4299, N-acetyl-alpha-glutamyl-phosphate reductase activity, inhibitor of granzyme A-activated DNase, AW124434, Histone H1(s), ME-IV, I-2PP1, Adenosine, dSET/TAF-Ibeta, 2610030F17Rik, S-Adenosylmethionine Sulfate Tosylate, TAF-IBETA, precursor ion, N-acetyl-aminoadipate semialdehyde dehydrogenase activity, Ion, quotient, Peptid, TAF-Ibeta, Polypeptide, assay, methyl cyanide, AA407739, i2pp2a, S Adenosylmethionine Sulfate Tosylate, S AdenosylmethionineBru, data, Radiation, human being, Raw, protein complex, number, high weight, Visible Light, Light, protein, protein-containing complex, CD258, Histone, presence, Peptide, Ly113, polypeptide, peptide, Histone H2b, Polypeptides, Histone H2a, protein polypeptide chains, count in organism, LIGHT, peptido, ClvPrd, native protein, natural protein, polypeptide chain, hnu, Protein, heavy, sequence, Del(8)44H, light, protein aggregate, foton, Histone H3.3, Data Base, Visible Radiations, Radiations, Svc, Visible Radiation, Col4a-1, HVEML, peptides, HVEM-L, Lichtquant, Photoradiation, proteins, Visible, man, Histone H1(s), primary structure of sequence macromolecule, human, LTg, light quantum, Photoradiations, Histone H5, photon, Histone H4, Histone H7, Peptid, peptidos, Polypeptide, TR2, quantitative, Peptide., Histone H1, Histone H3, gamma, presence or absence in organismandrogeno, Forms, IPP2A2, androgens, Par1c, Materials, insensitive, Lysine Hydrochloride, Metabolisms, neoplasia, Bioenergetics, carcinoma of the prostate, protein, prostate carcinoma., Tumor, androgene, DmelCG8201, 5730420M11Rik, DHTR, prostate gland carcinoma, Histone H2b, Histone H2a, protein polypeptide chains, Lysine Acetate, Oxidative, malignant tumor of prostate, AR, 2, Expenditure, protein aggregate, Chromatins, L Lysine, Gonadectomies, SET, Gene Expressions, reference sample, K, TAF-I, cell process disease, tumor disease, phosphatase 2A inhibitor I2PP2A, neoplasm (disease), malignant tumour of prostate, epsilon-diaminocaproic acid, proteins, TRMT11-2, DmelCG4299, set, IGAAD, dihydrotestosterone receptor, LYS, DmelCG10574, Lysin, signaling process, chromosome scaffold, Histone H5, Histone H4, xenotransplant, Histone H7, prostate gland cancer, tumor, CG16701, Histone H1, nuclear receptor subfamily 3 group C member 4, Histone H3, single organism signaling, Tumors, nuclear chromatin, phapii, cancer of the prostate, Prostates, Trm112p, CG8201, tumor of the prostate, familial, StF-IT-1, dMARK, neoplastic disease, CG11960, tumours, CG30131, CG30132, EMK, Androgen, DPAR-1, lysine, malignant neoplasm of prostate gland, Genetic Materials, NOS, tumour of the prostate, Genetic Material, malignant neoplasm of the prostate, malignant prostate tumor, Phosphorylation, HLA-DR-associated protein II, resistant, DI-2, I-2Dm, androgenes, alpha, CG4299, inhibitor of granzyme A-activated DNase, cancer of prostate, malignant tumour of the prostate, I-2PP1, tumour disease, malignant neoplasm of prostate, l(2)27C1, Energy Metabolisms, TAF-IBETA, BcDNA:RH48823, Material, Proliferation, malignant prostate tumour, MARK, Cistron, TAF-Ibeta, other neoplasm, i2pp2a, prostate, malignant tumor of the prostate, prostata, Proliferating, cytoplasmic chromatin, biological signaling, conformation, carcinoma of prostate gland, Neoplasms, Par-1c, Gene, malignant prostate gland neoplasm, protein-containing complex, NGP - new growth of prostate, PHAPII, NEOPL, HSPC170, malignant prostate neoplasm, template-activating factor I, polypeptide chain, dPAR-1, Metabolism, Lysine, dPar-1, neoplasm, prostate neoplasm, Histone H3.3, 27C1, Methyltransferase, Genetic, l(2)k06323, proliferating, ipp2a2, 2pp2a, Energy, tumour, Expenditures, respiratory-chain phosphorylation, Expressions, CG10574, carcinoma of prostate, PAR-1, MALIGNANT AND UNSPECIFIED (INCL CYSTS AND POLYPS), Oxidative Phosphorylations, 2PP2A, taf-ibeta, dSET, dSet, Par-1, Expression, HSPC152, relational structural quality, Methylations, Neoplastic Growth, Controlled, hereditary prostate cancer, par1, Controlling, data, prostatic neoplasm, prostatic cancer, protein complex, igaad, dPAR1, Phosphorylations, dPar1, 0610038D11Rik, Cistrons, Histone, androgenos, Cell, cancer of prostate gland, neoplastic growth, native protein, natural protein, I-2PP2A, Acetate, male prostate, Androgene, Dm I-2, Protein, anon-WO0210402.19, I2PP2A, methylation, L-Lysine, prostate carcinoma, Energy Expenditure, Gonadectomy, Castrations, Energy Expenditures, disease of cellular proliferation, Histone H1(s), TRM112, Bioenergetic, signalling, dSET/TAF-Ibeta, 2610030F17Rik, signalling process, DPar1, NR3C4, prostate cancer, PC, PAR1, Par1, 6-diaminohexanoic acid, AA407739, Enisyl, NEOPLASMS BENIGN, Neoplasiabiochemical pathways, Proliferating, Metabolic Process, Par1c, insensitive, carcinoma of prostate gland, Processes, Par-1c, Metabolic Concepts, carcinoma of the prostate, malignant prostate gland neoplasm, Metabolic Processes, NGP - new growth of prostate, DmelCG8201, prostate gland carcinoma, malignant prostate neoplasm, dPAR-1, Metabolism, malignant tumor of prostate, Concepts, Cell., Metabolism Concept, dPar-1, Phenomenon, Metabolism Phenomena, prostate neoplasm, Gonadectomies, 27C1, catabolism, l(2)k06323, proliferating, malignant tumour of prostate, Metabolic Concept, metabolic process resulting in cell growth, carcinoma of prostate, PAR-1, Par-1, biotransformation, prostate gland cancer, CG16701, Catabolism, hereditary prostate cancer, par1, cancer of the prostate, prostatic neoplasm, prostatic cancer, CG8201, tumor of the prostate, degradation, Process, metabolism resulting in cell growth, familial, dPAR1, dPar1, dMARK, CG11960, CG30131, Concept, Metabolic Phenomena, CG30132, EMK, Metabolism Concepts, cancer of prostate gland, anon-WO0210402.19, DPAR-1, Phenomena, malignant neoplasm of prostate gland, secretion, NOS, metabolism, tumour of the prostate, prostate carcinoma, Metabolic Phenomenon, Gonadectomy, malignant neoplasm of the prostate, malignant prostate tumor, multicellular organism metabolic process, Castrations, biodegradation, resistant, Metabolic, cancer of prostate, malignant tumour of the prostate, malignant neoplasm of prostate, l(2)27C1, single-organism metabolic process, BcDNA:RH48823, Proliferation, malignant prostate tumour, MARK, DPar1, prostate cancer, PC, PAR1, Par1, malignant tumor of the prostate, AnabolismtrueKMT9 writes the histone mark H4K12me1 and controls metabolism and proliferation of castration-resistant prostate cancer cellsPosttranslational modifications of histones such as methylation regulate chromatin structure and gene expression. Methylation of histone lysine residues is generally performed by SET domain methyltransferases. Here, we identify the heterodimeric C21orf127/TRMT112 complex as a specific histone methyltransferase. Assembly of the seven-b-strand protein C21orf127 (also named Hemk2, N6amt1 or PrmC) with TRMT112 is essential to form an active enzyme, hereafter named KMT9 that writes the histone mark H4K12me1 in vitro and in vivo. The H4K12me1 mark is enriched at promoters of KMT9 target genes and co-localises with the active histone mark H4K12ac. By controlling expression of genes involved in energy metabolism, KMT9 regulates oxidative phosphorylation in androgen receptor-dependent and -independent prostate tumour cells. Importantly, KMT9 depletion severely affects proliferation of castration and enzalutamide-resistant prostate cancer cells and xenograft tumours. Together, our data link the writing of the H4K12me1 histone mark by KMT9 with KMT9-dependent gene expression, which in consequence regulates energy metabolism and proliferation. KMT9 executes these functions independently of androgen receptor and androgen signalling thus, providing a promising paradigm for the treatment of castration resistant prostate cancer.2019-03-112018-02-15PXD008965646099295358511145284218809058113633363515242602758385NCBITaxon:1280688881114301187263555571145262364927467073641135461582158236889511232944222911809923233654583334607925193659143205971293865581914932637371250232NCBITaxon:12721119577481866584345135867221755437408169180454266834453058611496367445281132081003923751463694208964208963445974137979128384400667935293216599404832165972165954550131528221659132183217292042779447623764543172030949801975255569781439843948840479NCBITaxon:21575515343989903712285772059687623588845581817808173630943179698936160488409859NCBITaxon:3716210478215059073204913816936303555432046632633456545351217459216129410026420369456941016763410210761277351131658239254727933428146135843017463345774449167547604675684030191726518130194111929439411310239320253258278839347495165299835525663175854310243102459808757417555183201945268019083233115611415542193255521123598755529NCBITaxon:4751547641465477180066556484337677299767982510216998238976411557444474153NCBITaxon:5055710003646534933414754801946751299114359741276473137086326547876569835983819201198524794640986099586766815729513722117110110768783497419860148840633424923598598865597464998703920211107272151908503711708283502419027463604614737593925325664890507945265919722119151774208588596NCBITaxon:6157505210283079895110517237505957613172000263419610940772275689964975157244NCBITaxon:2NCBITaxon:170361250617476792745062724915961538400524643213686150596866629417347515363291018194301086731018563558633149286287132161865368595260710185687294110091416333NCBITaxon:619184023210007348824187878645159553470951451225786260704260705260707658457561007133705654195NCBITaxon:3055504457913998781681928752829126806315327341478281055524469008150251930300852NCBITaxon:5693826883064017946991510151548242837047592723708281677285035158199011951953370249240536088817562615835583337605726711935019980861193502167879371137127240737145712562745790311333550372847924910078711082NCBITaxon:961559463318586260799141262200298492670192812528303856959111006581237561275045426662551519788375036009412934973524721401257649908515151438992374537474454464152432359288705158059553632716029385114978633892894535936412754225117167387158955210103061296715845158493765327159100226376733889423451287689146479216413328038280353690989462181214108034817939228788929502711311749346819227321890382107601247190400727114311402064119810871355477142450722953379698129163470871415754699287770202060328114319338293550833291292211487252649997727550717629970767892683214053294128160783922737759620551693605330797255181135491597495350261159556143213840607219496340161433471046603815184547448500356709747751240607699468810376201488728909316435649408941442423277422140921157781154756929793244704749906604162749907312989913112273170187103589915103609833411307981336795316407103401578630333733013064139925306423654788103372994044586207340115357111952965261111471611313166343053120179935129938505731764261979401151345844577267872862386211607913922279835112552281167617573121080772861814084341033519963218496139248820963304393758626NCBITaxon:26970493398624369411031219588399758644864324995252972463438637256737863912334355141493179986865540553435291486545147124546610429863571246791516413588840193733450877182380391295297197486931028688133731054877125198507625547011938062563188657130344351655865886223864946454636493NCBITaxon:110711428006288356491845881087439145943854331116781077439334953469397738218993182269003512814595339966869595418694200308954486972443669545623355180426430869220030228481242642815047514991051029938626290351208955711700893062186441930615755841310165382352557722399463601063601043994754440423761021279056881311622962981767718709312811050231855796694654318632051974417394124501288531838891988531911137910371647351660NCBITaxon:10114284503163119857383432379560260871213463959842752959526643733432322354431427524224308143361959313345659432741699631122233605201216979869728643680121870733474740818829760381812498853401884397331274414163958885829554648038224326408172124193512816140559204254655194582283827127442340817019332981274420127442641399616601876254829647187695648188229148015412045513847384818829451906951166336036484981015106937352945297222585941097677191107927943447256848479387033384991722617561094616143512032581205136521850523525944732054944664499175582580388043330120678110806124619624619732063752299509068387911879473980071868015723074683574125811757150847388580329031388869152167783997844751295139870435126246969281239771333399510343048138182198131882682476292657163295719222219791169381979121247132838828139529491459541619341390363120416232644607112104117061274432108595669412831666943694559729410658330879478447517415080820654231744712702814111946691454587932920101981264690410661145755139251697316062121331283128212803395210832825253531522811235315349825741948147875362314241013567429510524261914548111506211292829965975319351630980069781036905660566156641624251187023482118705638632226186129986068851262321721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