Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/03/PXD012734/FVZ2397-S02-A-Sample_A_asynchron.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/03/PXD012734/FVZ2397-S03-A-Sample_B_mitotic.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/03/PXD012734/FVZ2397_txt.zipprimaryOK200johannes.gloeckner@dzne.deJohannes GloecknerMass SpectrometryShotgun proteomicsNot availableUsp9xUbiquitinylationhttp://www.ebi.ac.uk/pride/archive/projects/PXD012734Cell CultureFor SILAC-based ubiquitin-purification His-Ubiquitin over expression HEK 293T cells grown in either “heavy” (K-8/R-10) (USP9X shRNA) or “medium” (K-4/R-6) medium (control shRNA) were lysed in buffer containing 8M urea, 300mM NaCl, 0.5% NP40, 50mM Na2HPO4, 50mM Tris pH8.0, 20mM imidazole and proteasome inhibitors. For synchronization in mitosis HEK 293T cells were treated with nocodazole. The two samples were combined immediately after lysis using equal amounts of total protein to purify ubiquitinated proteins with Ni-NTA-Agarose. After 1.5h incubation, Ni-NTA-Agarose were washed once with lysis buffer, twice with Buffer A and twice with Buffer B (Buffer A: 8M urea, 300mM NaCl, 0.5% NP40, 50mM Na2HPO4, 50mM Tris pH (6.3); Buffer 1B: 8M urea, 300mM NaCl, 0.5% NP40, 50mM Na2HPO4, 50mM Tris pH (6.3), 10mM imidazole). Proteins were proteolytically cleaved with trypsin in 50mM Ammoniumbicarbonate pH 8.0 supplemented with 1% RapiGestTM directly on the Ni-NTA-Agarose, over night. Prior to proteolysis, Cysteine residues were reduced by DTT following carbamylation by Chloroacetamide. After proteolysis, the RapiGestTM surfactant was hydrolysed by TFA according to the manufacturer’s protocol and samples were subsequently desalted with StageTips (Thermo Fisher Scientific). Vacuum-dried samples were re-dissolved in 0.5% TFA and Extracted peptides were subsequently subjected to LC-MS/MS-analysis: Tryptic peptide mixtures were injected automatically and loaded at a flow rate of 30 μl/min in 0.1% trifluoroacetic acid in high performance liquid chromatography (HPLC)-grade water onto a nano trap column (300 μm inner diameter × 5 mm precolumn, packed with Acclaim PepMap100 C18, 5 μm, 100 Å; Thermo Scientific). After 3 min, peptides were eluted and separated on the analytical column (75 μm inner diameter × 25 cm, Acclaim PepMap RSLC C18, 2 μm, 100 Å; Thermo Scientific) by a linear gradient from 2% to 30% of buffer B (80% acetonitrile and 0.08% formic acid in HPLC-grade water) in buffer A (2% acetonitrile and 0.1% formic acid in HPLC-grade water) at a flow rate of 300 nl/min over 147 min. Remaining peptides were eluted by a short gradient from 33% or 30% to 95% buffer B in 5 or 10 min. Analysis of the eluted peptides was done on an Orbitrap Fusion Tribrid mass spectrometer (Thermo Fisher Scientific). From the high-resolution MS prescan in the Orbitrap with a mass range of 335 to 1,500 at a resolution of 120,000, at least doubly charged precursors were selected for fragment analysis with the top speed method and 3s cycle time. The normalized collision energy for higher-energy collisional dissociation (HCD) was set to a value of 30. The lock mass option was activated; the background signal with a mass of 445.12003 was used as lock mass. Every ion selected for fragmentation was excluded for 20 or 60 s by dynamic exclusion.Prideiodoacetamide derivatized residueSILACRAW files were analysed with MaxQuant (version 1.5.6.5) against the human subset of the Swiss-Prot database (release 2016_08) with the following settings: Lys-8/Arg-10 (referred as “heavy” label) and Lys-4/Arg-6 (referred as “medium” label) as isotopic pairs (multiplicity=2); Carbamidomethyl as fixed and Methionine oxidation and N-terminal acetylation as well as diGly (K) as variable modifications. The MaxQuant results were further processed with in house R-scripts for visualization and statistical analysis. Hits were reported as significantly enriched using the Significance B test at p=0.01 (Cox and Mann 2008). References: Cox, J., and M. Mann. 2008. 'MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification', Nat Biotechnol, 26: 1367-72.ProteomicsChristian Johannes GloecknerLTQ OrbitrapPARTIAL(1) German Center for Neurodegenerative Diseases (DZNE), 72076 Tübingen, Germany (2) University of Tübingen, Center for Ophthalmology, Institute for Ophthalmic Research, 72074 Tübingen, GermanyHomo Sapiens (human)32152317 Dietachmayr M, Rathakrishnan A, Karpiuk O, von Zweydorf F, Engleitner T, Fernández-Sáiz V, Schenk P, Ueffing M, Rad R, Eilers M, Gloeckner CJ, Clemm von Hohenberg K, Bassermann F. Antagonistic activities of CDC14B and CDK1 on USP9X regulate WT1-dependent mitotic transcription and survival. Nat Commun. 2020 11(1):1268 10.1038/s41467-020-15059-5johannes.gloeckner@dzne.deBiologicalDZNEGermanyRegulation of mitosis secures cellular integrity and its failure critically contributes to the development, maintenance, and treatment resistance of cancer. In yeast, the dual phosphatase Cdc14 controls mitotic progression by antagonizing Cdk1-mediated protein phosphorylation. By contrast, specific mitotic functions of the mammalian Cdc14 orthologue CDC14B have remained largely elusive. Here, we find that CDC14B antagonizes CDK1-mediated activating mitotic phosphorylation of the deubiquitinase USP9X at serine residue 2563, which we show to be essential for USP9X to mediate mitotic survival. Starting from an unbiased proteome-wide screening approach, we specify Wilms' tumor protein 1 (WT1) as the relevant substrate that becomes deubiquitylated and stabilized by serine 2563-phosphorylated USP9X in mitosis. We further demonstrate that WT1 functions as a mitotic transcription factor and specify CXCL8/IL-8 as a target gene of WT1 that conveys mitotic survival. Together, we describe a ubiquitin-dependent signaling pathway that directs a mitosis-specific transcription program to regulate mitotic survival.Antagonistic activities of CDC14B and CDK1 on USP9X regulate WT1-dependent mitotic transcription and survival.Dietachmayr Michael M, Rathakrishnan Abirami A, Karpiuk Oleksandra O, von Zweydorf Felix F, Engleitner Thomas T, Fernández-Sáiz Vanesa V, Schenk Petra P, Ueffing Marius M, Rad Roland R, Eilers Martin M, Gloeckner Christian Johannes CJ, Clemm von Hohenberg Katharina K, Bassermann Florian FR 17934, sodium salt, Eph-like tyrosine kinase 1, (5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl)-, ammonium formate, 13C-labeled, MeCN, cadmium salt, Proteasome Inhibitor, PLXN4, ion, PLXN3, fond, Phases, Basodexan, magnesium formate, CycEI, zinc salt, Long Term, 5730420M11Rik, Trifluoroacetate, Polypeptides, protein polypeptide chains, cobalt(II) formate dihydrate, CH3-C#N, B1, imidazole monohydrochloride, Urea, Human embryo kinase, Ubiquitinated, TYRO4, WMS, RCB2202, peptidolysis, HEK293T, nocodazol, SET, Carbamic acid, Divorced, M Phases, EK4, phosphatase 2A inhibitor I2PP2A, proteins, Divorces, 934, HPLC, AI047805, DmelCG4299, AA407302, salt, set, C1 4B, decreased, methyl [5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl]carbamate, column, 6-anhydro-alpha-L-galactopyranosyl-(1->3)-beta-D-galactopyranan, D1, n, autolysin activity, Half Cystine, 6.3, nickel salt, SGS, LC-MS2, Degradations, Zinc Cysteinate, cobalt (+2) salt, Liquid Chromatography, Longterm Effect, Cyc E, CEP52, ACMICD, common salt, sodium (4:1:1) salt, BG:DS07108.3, Dm1, magnesium salt, [4)-3, Sepharose C1, l(2)05206, Ubiq, Hydrogen Oxide, AA407699, Acid, HLA-DR-associated protein II, Mitoses, DI-2, I-2Dm, nocodazole, Protein Digestions, formate, cromium (+3), inhibitor of granzyme A-activated DNase, study protocol, beta Trypsin, I-2PP1, TAF-IBETA, liquid chromatography-tandem mass spectroscopy, bacteriolytic toxin activity, ammonium (4:1) salt, Mitotic, TAF-Ibeta, methyl cyanide, lead (+2) salt, nickel formate dihydrate, cycline, lead formate, Effects, DmcyclinE, aluminum salt, ATP Dependent Proteolysis Factor 1, Carbamide, protein-containing complex, CG7826, Human, cdi7, period, cyclinE, LC-MSMS, Gene Products, CG7835, Cdi7, CDI7, AL022749, CG42273, H2im(+), Half-Cystine, Carmol, Cesium Trifluoroacetate, iones., RGD1560056, thallium (+1) salt, Longterm, L Cysteine, beta-Trypsin, rubidium salt, 2pp2a, Long-Term, methanoic acid, DmcycE, ions, CG10574, 40S ribosomal protein S27a, 2PP2A, HEK-293T, Kochsalz, dSET, dSet, peptidos, protein tagging activity, methyl N-(5-thenoyl-2-benzimidazolyl)carbamate, ETK, Al-imidazole, PTHB1, 1-(14)C-labeled, Proteins, AL022658, potassium formate, copper (+2) salt, acetonitrile, buffer, Digestions, Cell, polypeptide, Ubiquitin A-52 residue ribosomal protein fusion product 1, l(2)k02514, native protein, DmCycE, Natriumchlorid, Proteolyses, carbonyldiamide, I-2PP2A, Ubiquitin-related 2, C18, cupric formate, chemical analysis, Long Term Effects, Dm I-2, Mitotic M Phase, MFS1, background, NaCl, aluminum formate, AU020952, covalent modifier, Phase, lithium formate, holin, l(2)k02602, underdeveloped, Separations, MRX99, ethanenitrile, imidazolium, (1->4)-3, R17934, Longterm Effects, ME-IV, plan specification, NSC 238159, Gene Proteins, D-CycE, 60S ribosomal protein L40, protein tag, liquid chromatography tandem mass spectrometry, Ubiquitin-related, R17, liquid chromatography tandem mass spectroscopy, imidazole acetate, IPP2A2, NCMe, determination, Sepharose C1 4B, FAF-X, zinc formate, ETK1, lead salt, protein, lysis, peptide, Mitotic M, halite, ammonium tetraformate, peptido, ClvPrd, Ccne, Inhibitors, Min, protein aggregate, present in fewer numbers in organism, Effect, DmelCG42273, Sepharose, LCMSMS, High Mobility Protein 20, reference sample, peptides, ATP-dependent proteolysis, TAF-I, E927b, hypoplasia, nocodazolum, min, Ubiquitin-related 1, mAPC, copper, Protein Degradation, Hek 293T, cloruro sodico, APF-1, IGAAD, lithium salt, DmelCG10574, R-17934, HSSEXGENE, trifluoro-, necrosis, GPHYSD2, ammonium (2:1) salt, Long-Term Effects, phapii, rock salt, XAP-6, 4B, LC-MS/MS, Dmel_CG7826, StF-IT-1, DFFRX, br37, Thermo Scientific, ur, shortened, Dffrx, carbamide, imidazole conjugate monoacid, HCD, Ionen, Digestion, (5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl)-carbamic acid methyl ester, EPH-like kinase 4, bacteriocin activity, nickel (+2) salt, Dmel_CG7835, Acetic acid, Ubiquitin, Fafl, Mnb, MNB, ammonium salt, Trifluoroacetic, Human Ubiquitin, cobaltous formate, hEK4, L-Cysteine, MASS, CG4299, AW124434, natrii chloridum, 293 T, surfactant, chloracetamide, H2NC(O)NH2, NSC238159, imidazolium chloride, lysin activity, l35Dd, Cesium, FAF, oncodazole, surface active agent, copper salt, HEK, Sepharose 4B, FAM, imidazolium ion, Polypeptide, Tyrosine-protein kinase receptor ETK1, short, Inhibitor, i2pp2a, 1728, time, IMIDAZOLE, N-(5-(2-thienoyl)-2-benzimidazolyl)carbamic acid methyl ester, Cysteine Hydrochloride, 293tsA1609neo, Degradation, cesium salt, Proteasome Endopeptidase Complex Inhibitors, surfactants, FBN, Gene, cycE, Ubiquitin carboxyl extension protein 80, l(2)br37, PHAPII, 293T, LC-MS-MS, CYCLE, Buffer, formic acid, Karbamid, method, potassium salt, HEK4, polypeptide chain, reduced, template-activating factor I, subnumerary, ECTOL1, 14C-labeled, method used in an experiment, Harnstoff, imidazole citrate, HMG-20, tiny, ubiquitin-like protein modifier, Separated, Tyrosine-protein kinase TYRO4, CYCE, 293-T, DmelCG3938, CyclE, ipp2a2, chlorure de sodium, stubby, 3938, decreased number, calcium formate, Protein Degradations, OCTD, l(2)k05007, dm-cycE, 2.7.10.1, taf-ibeta, ubiquitin, Agarose, Cu-imidazole, cromium (+3) salt, Long-Term Effect, imidazole sodium, 6-An-alpha-L-Galp-(1->3)-beta-D-Galp-(1->]n, sodium formate, Controlled, imidazole monophosphonate, small, Human Embryonic Kidney 293T, Controlling, DYRK1, nickel formate, 3H-labeled, protein complex, igaad, methyl ester, strontium formate, HEK 293 T, N-(5-(2-thenoyl)-2-benzimidazolyl)carbamic acid methyl ester, Protein Digestion, strontium salt, Peptide, Proteasome, NSC-238159, methyl (5-(2-thienylcarbonyl))-1H-benzimidazole-2-ylcarbamate, LC/MS/MS, natural protein, mitosis, Protein, I2PP2A, table salt, Dyrk1, Vacuums, CyeE, ATP-Dependent Proteolysis Factor 1, 1H-imidazol-3-ium, M Phase, ACETONITRILE, cyanomethane, WMS2, Separation, 10^[-9], uree, urea, CC1, l(2)35Dd, Tripcellim, chromic formate, UREA, introduction, Protein Gene Products, Trypure, dSET/TAF-Ibeta, 2610030F17Rik, Ion, velocity, SSKS, Mitotic M Phases, calcium salt, Peptid, assay, Oncodazole, AA407739, CG3938, HEK-293-T, 5730589N07Rik, sodium chlorideSRM5, hereditary kidney adenosarcoma, type 1, transcription, 2.7.11.22, familial Wilms tumour, WIT-2, hereditary Wilms' tumor, AWT1, 2.7.11.23, CDC14B, FAF-X, GUD, autosomal dominant, WAGR, hCDC14B, Cell division control protein 2 homolog, EWS-WT1, cdc, familial Wilms' tumor, cdc14beta, Cdc2a, l(2)31Eh, swo2, AL022749, Wilms' tumour, Wilms tumor 1, xcdc14b, Dmcdc2, xwt1, XWT1a, RGD1560056, XWT1b, CDC28A, xWT1, AA472821, P34CDC2, hereditary nephroblastoma, renal Wilms tumour, Wilms tumor type 1, A530086E13Rik, Wilms tumour type 1, AA407302, DNA-dependent, CDK1/CDC2, cdc2Dm, 2810432N10Rik, cdk1, CG5363, dCdk1, Wilms' tumor, Cdc2, CDC2, ZFF29, wt-1, CDK1, Dcdc2, Cdk1, transcription from bacterial-type RNA polymerase promoter, AFF29, somatic mutation, Dm cdc2, hereditary Wilms tumor, CDC14B3, familial Wilms' tumour, cellular transcription, DNA-dependent transcription, AL022658, HSL5, DFFRX, familial Wilms tumor, Wilms tumor, somatic, renal Wilms tumor, Dffrx, survival, group 4, DmCdc2, hereditary renal adenosarcoma, DmCdk1, DmelCG5363, p34-lt-CDC2-gt-, AA407699, Fafl, XeWT1, 5363, MRX99, nephroblastoma, DNA-templated, WT33, tws1, hereditary Wilms' tumour, Cell division protein kinase 1, p34<CDC2>, Cdc14B1, Cdc14B2, p34 protein kinase, FAF, bacterial transcription, time of survival., Cdk-1, FAM, wt1, WT1, NPHS4, CDCDm, 5730589N07RikNAT, Nat, Bru, 1110033A15Rik, cox4, 11alpha, human being, 2-amino-4-(methylsulfanyl)butanoic acid, D17H6S113E, alpha-L-Talopyranuronic acid, Raw, TalANAc, L-arginine, AI326280, L Isomer, FBN, alpha-L-N-acetyltalosaminuronic acid, protein, broad, DNB1, protein-containing complex, FSH, L-(+)-arginine, peptide, CG10664, Polypeptides, L-Isomer Methionine, protein polypeptide chains, l(3)rQ526, Methionine, ClvPrd, peptido, (2S)-2-amino-5-(carbamimidamido)pentanoic acid, polypeptide chain, ECTOL1, IV, L-Methionine, 2, Del(8)44H, ARG, Arg, DmelCG10664, protein aggregate, sty, l(3)j5E11, Pedameth, WMS, ARP, Svc, DmelCG18104, mKIAA0458, (S)-2-amino-5-guanidinopentanoic acid, Coproporphyrinogenase, peptides, K, Gm17754, COXIV, ATN1L, M, (S)-2-Amino-5-guanidinovaleric acid, R, epsilon-diaminocaproic acid, Argos, Acetylations, AW742570, CG4531, proteins, arg, Ring3, man, COXA_DROME, OCTD, 13E, Lys, LYS, 2-amino-4-(methylthio)butanoic acid, Cox4, Lysin, DmAos, COX, L-Arg, peptidos, 1.3.3.3, metionina, GPHYSD2, Racemethionine, COX-VA, rlt, statistical analysis, Lzm, SGS, ABLL, L-Arginin, Lyzs, visualization, Fsrg1., Lzp, eye-lt-m03Jus-gt-, wide/broad, DL-Methionine, Atr2, protein complex, acetylation, EG:171D11.4, 2-Amino-4-(methylthio)butyric acid, arginine, total expressed protein, L-Isomer, 2-(acetylamino)-2-deoxy-, results, Peptide, Abll, ACMICD, polypeptide, l(3)rJ812, RING3, native protein, natural protein, VA, AL024441, AW228947, Protein, FSRG1, lysine, Methionin, IV-1, MFS1, Lzm-s1, methionine, mKIAA4005, Hmet, COX IV, l(3)05959, fixed, AI414665, CG18104, Data Base, Met, WMS2, L-Arginine, D6S113E, Col4a-1, gil, Va, Coprogen oxidase, COX IV-1, Rnf3, RNF3, (2S)-2-amino-5-guanidinopentanoic acid, Frg-1, tend, eyes3, alpha, MASS, 15S)-, human, l(3)05845, DmelCG4531, Lysm, data encoding as image, Aos, wide, CG14724, (5Z, l(3)rJ472, SSKS, alpha-amino-gamma-methylmercaptobutyric acid, AA536808, Peptid, DmelCG14724, Polypeptide, EG:65F1.3, variable, 6-diaminohexanoic acid, Liquimeth, Proteomes, Fsrg-1, alpha-L-2-N-acetylamino-2-desoxytaluronic acid, Cox4aEph-like tyrosine kinase 1, thiol-dependent ubiquitinyl hydrolase activity, 293tsA1609neo, ubiquitinyl hydrolase 1 activity, FAF-X, CDC14B, Aminosaeure, Aminocarbonsaeure, Amino acid, Phases, Gene, ETK1, ATP Dependent Proteolysis Factor 1, alpha-amino acid, Ubiquitin carboxyl extension protein 80, Isotope-Coded Affinity, deubiquitylase, thiol-dependent ubiquitin-specific protease activity, hCDC14B, 293T, Human, Mitotic M, deubiquitinase activity, HEK4, cdc14beta, Gene Products, AL022749, HMG-20, Isotopically-Coded Affinity, ubiquitin-like protein modifier, Human embryo kinase, TYRO4, Isotopically-Coded Affinity Tagging, Tyrosine-protein kinase TYRO4, RCB2202, xcdc14b, HEK293T, ubiquitin-specific protease activity, ubiquitin hydrolase activity, RGD1560056, amino acids, M Phases, 293-T, High Mobility Protein 20, reference sample, EK4, AA472821, Ubiquitin-related 1, proteins, ubiquitin C-terminal hydrolase, A530086E13Rik, AA407302, Hek 293T, UBP, APF-1, 40S ribosomal protein S27a, 2.7.10.1, HEK-293T, 2810432N10Rik, Stable, ubiquitin, Isotope-Coded Affinity Tagging, ZFF29, Tagging, protein tagging activity, AFF29, Stable Isotope Labeling, thiol-dependent deubiquitinase, ETK, Controlled, ubiquitin C-terminal hydrolase activity, ubiquitinyl hydrolase activity, Human Embryonic Kidney 293T, CDC14B3, Controlling, Aminokarbonsaeure, Proteins, AL022658, HEK 293 T, DFFRX, alpha-amino acids, CEP52, Labeling, alpha-amino carboxylic acids, Cell, Isotope, Dffrx, Ubiquitin A-52 residue ribosomal protein fusion product 1, Stable Isotope, Ubiquitin-related 2, mitosis, Protein, EPH-like kinase 4, Mitotic M Phase, ATP-Dependent Proteolysis Factor 1, M Phase, Isotope Coded Affinity Tagging, Ubiq, AA407699, Fafl, Ubiquitin, Amino Acid, Acid, covalent modifier, Phase, Amino acids, Mitoses, Protein., MRX99, Human Ubiquitin, hEK4, Isotope Labeling, Amino, Affinity Tagging, 293 T, Isotopically-Coded, UCH2, Protein Gene Products, Gene Proteins, Cdc14B1, deubiquitinating enzyme, Cdc14B2, 60S ribosomal protein L40, FAF, protein tag, deubiquitinase, HEK, Mitotic M Phases, Isotope-Coded, FAM, Ubiquitin-related, Acids, Mitotic, Tyrosine-protein kinase receptor ETK1, HEK-293-T, 5730589N07Rikmalignant Growth, SRM5, transcription factor, type 1, T-cell leukemia, thiol-dependent ubiquitinyl hydrolase activity, target gene, LYNAP, single-organism developmental process, 2.7.11.22, embryonal nephroma, Monocyte-derived neutrophil chemotactic factor, WIT-2, 2.7.11.23, FAF-X, AGL4, Phases, protein, regulation of mitosis, Neutrophil-activating factor, IL-8(6-77), phosphorylation, thiol-dependent ubiquitin-specific protease activity, hCDC14B, 2-Amino-3-hydroxypropionic acid, Cell division control protein 2 homolog, EWS-WT1, cdc, protein polypeptide chains, MDNCF-c, familial Wilms' tumor, Mitotic M, Lymphocyte-derived neutrophil-activating factor, IL-8, RNA polymerase II distal enhancer sequence-specific DNA binding transcription factor activity, Cdc2a, l(2)31Eh, protein aggregate, IL8|NAP1 form III, MDNCF-a, MDNCF-b, signal transduction by protein phosphorylation, Dmcdc2, (Ala-IL-8)77, ubiquitin hydrolase activity, CDC14a1, M Phases, High Mobility Protein 20, neoplasm (disease), F10N7_150, AA472821, hereditary nephroblastoma, Ubiquitin-related 1, proteins, Wilms tumor type 1, ubiquitin C-terminal hydrolase, RNA polymerase II core promoter proximal region sequence-specific DNA binding transcription factor activity, IL8|NAP1 form IV, CA, AA407302, APF-1, DNA-dependent, CDC14A2, cdc14, cdc2Dm, MDNCF, kidney Wilms tumor, 2-amino-3-hydroxypropanoic acid, CG5363, 2810432N10Rik, malignant neoplasm, dCdk1, IL8|NAP1 form II, Cdc2, CDC2, AGAMOUS-like 4, zinc ion regulated proximal promoter sequence-specific DNA binding, CDC14, sequence-specific transcription regulatory region DNA binding RNA polymerase II transcription factor recruiting transcription factor activity, GCP|IL-8 protein V, MONAP, transcription from bacterial-type RNA polymerase promoter, phosphatase, Cdc14, GCP|IL-8 protein I, somatic mutation, thiol-dependent deubiquitinase, TRANSCRIPTION FACTOR, Dm cdc2, ubiquitin C-terminal hydrolase activity, RNA polymerase II distal enhancer sequence-specific binding, Deubiquitinating, RNA polymerase II core promoter proximal region sequence-specific binding, Emoctakin, ubiquitinyl hydrolase activity, malignancy, CDC14B3, GCP|IL-8 protein III, wide/broad, functional failure, DNA-dependent transcription, IL-8(7-77), monocyte-derived neutrophil-activating peptide, Wilms' tumour of the kidney, signal transduction by conformational transition, DFFRX, renal Wilms' tumour, familial Wilms tumor, T-cell chemotactic factor, somatic, metal ion regulated sequence-specific DNA binding RNA polymerase II transcription factor activity, CEP52, Granulocyte chemotactic protein 1, signal transduction by trans-phosphorylation, Programs, Dffrx, signaling cascade, childhood renal Wilms tumour, adult renal Wilms' tumour, 3-Hydroxyalanine, organ system cancer, DmCdc2, failure, sequence-specific distal enhancer binding RNA polymerase II transcription factor activity, DmelCG5363, Ubiq, AA407699, Fafl, Ubiquitin, F14P3.4, Yeast, nonanaplastic renal Wilm's tumour, Serine, GCP|IL-8 protein IV, Wilms tumor of the kidney, renal Wilms' tumor, Mitoses, Deubiquitinases, Interleukin-8, F14P3_4, Human Ubiquitin, nephroblastoma, IL8|NAP1 form VI, childhood renal Wilms' cancer, GCP|IL-8 protein II, Transcription factor, childhood renal Wilms tumor, Neutrophil-activating protein 1, renal embryonic tumor, Cdc14B1, wide, Cdc14B2, signaling pathway, p34 protein kinase, FAF, bacterial transcription, copper ion regulated core promoter proximal region sequence-specific DNA binding RNA polymerase II transcription factor activity, NAP1, Cdk-1, NAF, FAM, L Serine, Mitotic, WT1, wt1, L-Serine, cancer, metal ion regulated sequence-specific DNA binding, Proteomes, adult nephroblastoma, metal ion regulated proximal promoter sequence-specific DNA binding, hereditary kidney adenosarcoma, transcription, NAP-1, familial Wilms tumour, hereditary Wilms' tumor, AWT1, cell type cancer, ubiquitinyl hydrolase 1 activity, CDC14B, ATP Dependent Proteolysis Factor 1, Deubiquitinating Enzyme, GUD, disfunctional, broad, SEPALLATA 2, Ubiquitin carboxyl extension protein 80, protein-containing complex, monocyte-derived neutrophil chemotactic factor, autosomal dominant, deubiquitylase, WAGR, Human, protein amino acid phosphorylation, deubiquitinase activity, zinc ion regulated core promoter proximal region sequence-specific DNA binding, cdc14beta, polypeptide chain, protein 3-10C, Protein 3-10C, resistance, IL-8(1-77), Wilms tumour (nephroblastoma), swo2, AL022749, HMG-20, Wilms' tumour, Wilms tumor 1, sequence-specific DNA binding, ubiquitin-like protein modifier, neoplasm, IL-8(8-77), malignant tumour, xcdc14b, nonanaplastic renal Wilm's tumor, ubiquitin-specific protease activity, xwt1, XWT1a, RGD1560056, XWT1b, CG7134, GCP|IL-8 protein VI, CDC28A, xWT1, P34CDC2, Wilms tumour of the kidney, renal Wilms tumour, Maintenances, A530086E13Rik, Wilms tumour type 1, IL-8(5-77), UBP, Chemokine (C-X-C motif) ligand 8, 40S ribosomal protein S27a, signalling pathway, Enzyme, CDK1/CDC2, cdk1, RNA polymerase II proximal promoter sequence-specific DNA binding, time of survival, Wilms' tumor, copper ion regulated proximal promoter sequence-specific DNA binding, ubiquitin, renal embryonic tumour, signal transduction by cis-phosphorylation, ZFF29, IL8, emoctakin, hCDC14, granulocyte chemotactic protein 1, wt-1, CDK1, Dcdc2, Cdk1, protein tagging activity, AFF29, hereditary Wilms tumor, serine, CXCL8, familial Wilms' tumour, cellular transcription, LECT, protein complex, malignant, GCP-1, Phosphorylations, AL022658, total expressed protein, HSL5, Deubiquitinase, Wilms tumor (nephroblastoma), Wilms' tumor of the kidney, zinc ion regulated core promoter proximal region sequence-specific DNA binding RNA polymerase II transcription factor activity, homeobox 1, defective, Wilms tumor, LUCT, Monocyte-derived neutrophil-activating peptide, (Ser-IL-8)72, GCP1, Enzymes, transcription factor activity, development, renal Wilms tumor, Serin, IL8|NAP1 form I, Ubiquitin A-52 residue ribosomal protein fusion product 1, MT, survival, native protein, natural protein, IL8|NAP1 form V, Ubiquitin-related 2, mitosis, malignant neoplasm (disease), group 4, Protein, RNA polymerase II transcription factor activity, hereditary renal adenosarcoma, Mitotic M Phase, IL-8(9-77), ATP-Dependent Proteolysis Factor 1, signalling cascade, DmCdk1, M Phase, metal ion regulated core promoter proximal region sequence-specific DNA binding RNA polymerase II transcription factor activity, KNOTTED1-like homeobox gene 5, p34-lt-CDC2-gt-, XeWT1, covalent modifier, 5363, Phase, primary cancer, interleukin-8 receptor ligand, MRX99, Wilms tumour, DNA-templated, phosphoric monoester hydrolase activity, WT33, tws1, sequence-specific DNA binding RNA polymerase II transcription factor activity, neutrophil-activating protein 1, hereditary Wilms' tumour, metal ion regulated core promoter proximal region sequence-specific binding, copper ion regulated core promoter proximal region sequence-specific binding, OAF3, malignant tumor, DmelCG7134, Cell division protein kinase 1, F10N7.150, UCH2, p34<CDC2>, deubiquitinating enzyme, C-X-C motif chemokine 8, 60S ribosomal protein L40, malignant neoplastic disease, protein tag, time of survival., A830059A17Rik, deubiquitinase, Mitotic M Phases, Ubiquitin-related, NPHS4, CDCDm, CDC-14, 5730589N07Rik, sequence-specific transcription regulatory region DNA bindingAA407699, Fafl, AA407302, Dffrx, RGD1560056, FAF, FAF-X, MRX99, AL022749, AL022658, FAM, DFFRX, 5730589N07Rik.falseIdentification of mitotic substrates of phospho-regulated USP9XWe previously identified the deubiquitylase USP9X as a G2/M-specific CDC14B interactor (PXD012732) and subsequently identified USP9X as a specific CDC14B target (PXD012733). To investigate relevant mitotic substrates of phospho-regulated USP9X, we next performed a Stable isotope labeling by amino acids in cell culture (SILAC)-based screen in which ubiquitylated proteins were purified from control or USP9X depleted HEK 293T cells that were either asynchronous or synchronized in mitosis. Stable expression of recombinant His-tagged ubiquitin has been used for selective enrichment of ubiquitinylated proteins in both conditions.2020-03-252019-02-15PXD0127342806551646099180498511145422492156843544071284218327042363336351198343006636362758385696363121766668888111430155571145263145262364934613126957366649239161936413645106656889511363647627236611123294422295558326813042323122586365434607925193659106321432059712938655819125023254537111957748186619682158672256807617554373592536685862740816918045427877512010892250583674147956410039236942049433445974142001258641935293NCBITaxon:776436715675936438677160540483131528269304562716391624292042779447623763696121810032962922113763948840479NCBITaxon:2157131527310510671250278205968720165163684071736309698936104782148364320429294101061320491381693743277320464535111103217459216129288671288676410069456941016763410210761277351131658213584301417298174633675412873567568403013506218130194111929439367110411364459139498432025301270320221498751104759412047886134749515372110971652998279051355513980317585439801260866398081375755518320196447111415542193255521138343911931819813981810269701069955529NCBITaxon:47518502983354764146547798251021691572739823897648407413735186287054824153NCBITaxon:50557984467525100036433414754802103597413708115466547898353221599838106771034819852348780422531488414222863226634794615718712593991599191572952929211752719860148840633424951869123423523815233598589859886559746499870392021110723635272159874190828743418235024145113969293190259390719017463601237474636172262536001319722119158531058859698951105198997237989772000263419610264266722758858156899628737562690724420237172019998570724074767927459615333554319311671833554184005136861505968666211961748666134751536329NCBITaxon:176328628711033348837670580271848186536859598607575527472036105135294296110098402321000734882412336363071047005187878945556449447NCBITaxon:13356262664955419591399878168119011478281055524NCBITaxon:960613585846900813668481979153519307285930085264114090051184995262254121337591390143811139442370416744312044143708351581990119519532590841370137023943222774862615835583715371633376057267998086371137121906503714759172073127262981274575727090309031637910372814243371421044792491108239412NCBITaxon:961565885814780269266100658137352750454135663816475203750644223669502129349790546322455525365199764990814389923745374726473237602647312887055955361416915252740327160293851149786479436375529382479437225117752991204474167387217509193844137659393432715910022637674234442345467001287689591198467041464792936528034280382803510803481793921126212295027113118037386819112652699710286225273910760159736114324250711409810871355477436581424507229533796982042321242233218033698223470816457331858897165467599287770203829314125295508114817448886437452649997283035718308550717629970767892683704151562887651388563924561152086595551655181135491432137159749535026143213858781707683194963401614280053164686275110466038151845474485009747697477118051240174841855601555479204274201488711053253297261005941289093164351211604990013894382835423242331082934122058560416299131122731701879915429131267818187104931640729946714423683033373304852646383011306413992530642369923121162065261610337286129940207340115357652611114716133865431201725323712552329935316317NCBITaxon:1108499384106326787286232193348620862116079139222768119612552281495393213223233477412182753291757312994886181408434103351996313924891392488115338115339330439375569928626559933911209488624157183997586444237486435332444260665332413439097682127178637197985686399983998686553529148654104298635735825763288401986498238011191102868899958665220341632138119380625631886571303443112130886581144889198466645463NCBITaxon:11072NCBITaxon:11071142800628835374375116597145943116598854331116781077854314408043933434NCBITaxon:11082693977381382189931824431935128387145953116885311213278695berry8694100333586972443664701501114286921504751499105484204714722354025120847014649744497431170089306218644193061107632330805660396016217974825853946055396513212790568813377512981766017112147861050236694657349632051124501897441739412450128853183889198679488531911137910371647351660152943631631198573896939343237956026087121346334323223544314275241433611334565560291699636436802790105603140818838185733703401881274414437552196969716203871398558408172405598609183827275921274423408170275981274420127442623545513656533821176275943282474801199150956481882291690255120455148167238473848190695148498101510693731263753405203860186497405224052141856451804NCBITaxon:81347800915227548479387033384938719172261756109461614351203258120550523525944732054938641125722651216512538803766861206781122022307432201186705118670573873348691219387918680239800718680121545146835140818615084712067776918388516086113888691521677867212790100870435123171969281239294867713315484542261262198131882682411229265716329576938124712461485451328388348392949113903631204162665890771514880036942694169436945187901015080817053942065421055687317447127015329242814111946691169601454586953201019812646905505371408157697392830312383911283128212802811010832825253531522811235315349825741948153623142410145481N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