{"database":"MassIVE","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Other":["ftp://massive-ftp.ucsd.edu/v03/MSV000086339/"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":{"citationCount":0,"reanalysisCount":0,"viewCount":0,"searchCount":0},"additional":{"submitter":["Laurence Florens"],"full_dataset_link":["https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=0de7683d5b2945989b5ae5bb2fcb818f"],"submitter_email":["laf@stowers.org"],"sample_protocol":[""],"repository":["MassIVE"],"file_size":["67"],"ptm_modification":["MOD:01060 - \"A protein modification that effectively converts an L-cysteine residue to S-carboxamidomethyl-L-cysteine.\"","MOD:00719 - \"A protein modification that oxygenates an L-methionine residue to one of the diastereomeric L-methionine sulfoxide residues.\""],"data_protocol":[""],"omics_type":["Proteomics"],"instrument_platform":["Orbitrap Fusion Lumos"],"species":["Saccharomyces Cerevisiae (ncbitaxon:4932)"],"submitter_affiliation":["The Stowers Institute for Medical Research"],"pubmed_abstract":["The ring-like cohesin complex plays an essential role in chromosome segregation, organization, and double-strand break repair through its ability to bring two DNA double helices together. Scc2 (NIPBL in humans) together with Scc4 functions as the loader of cohesin onto chromosomes. Chromatin adapters such as the RSC complex facilitate the localization of the Scc2-Scc4 cohesin loader. Here, we identify a broad range of Scc2-chromatin protein interactions that are evolutionarily conserved and reveal a role for one complex, Mediator, in the recruitment of the cohesin loader. We identified budding yeast Med14, a subunit of the Mediator complex, as a high copy suppressor of poor growth in Scc2 mutant strains. Physical and genetic interactions between Scc2 and Mediator are functionally substantiated in direct recruitment and cohesion assays. Depletion of Med14 results in defective sister chromatid cohesion and the decreased binding of Scc2 at RNA Pol II-transcribed genes. Previous work has suggested that Mediator, Nipbl, and cohesin connect enhancers and promoters of active mammalian genes. Our studies suggest an evolutionarily conserved fundamental role for Mediator in the direct recruitment of Scc2 to RNA Pol II-transcribed genes."],"pubmed_title":["Mediator recruits the cohesin loader Scc2 to RNA Pol II-transcribed genes and promotes sister chromatid cohesion."],"pubmed_authors":["Mattingly Mark M, Seidel Chris C, Muñoz Sofía S, Hao Yan Y, Zhang Ying Y, Wen Zhihui Z, Florens Laurence L, Uhlmann Frank F, Gerton Jennifer L JL"],"pubmed_title_synonyms":["rpII140/wimp, Materials, l(3)Z23, RNA pol II, 8WG16, Pol-II, Gene, CG1163, RpII215, RpII140[wimp], nipbl, II, IDN3, Pol II, PolIIc, RNA polII, l(1)L5, RNApol2, POL, RNA pol IIo, Pol IIo[Ser2], 5, Cohesin, l(3)RplII140, RNApolII, Genetic, scc2, RNAP, Pol II CTD, Rpll140, Rpll18, RPII215, RNAPII0, BcDNA:RH21608, CDLS1, Ubl, EMBRYO DEFECTIVE 2773, T20K14_150, RNAP II, DNA Dependent RNA Polymerase II, L5, CG3180, l(1)DF912, RNA Pol II CTD, polII, CG1554, SISTER-CHROMATID COHESION 2, Pol IIo[ser5], Pol II0[ser2], l(1)G0040, RNA PolII, Pol IIo, RNA Pol II 140, Pol IIc, Pol IIa, l(1)DC912, PolII, Cohesin Protein Complex, Cistrons, RNAPII, DmelCG1163, RPII140, RPB2_DROME, CG3284, Dm6, l(3)88Be, RNA Pol IIc, Genetic Materials, DmRP140, CDLS, Scc2, DmelCG3180, SCC2, Rpll215, scc2-2, Genetic Material, scc2-1, pol II, IDN3-B, Pol-IIa, wimp, RPB6_DROME, ARABIDOPSIS THALIANA SISTER-CHROMATID COHESION 2, DmelCG1554, RP140, T20K14.150, Rpb2, Rpb1, RNA Pol II, RPB1, Pol IIo[ser2], PolIIo, RNAP II LS, RpABC14, RpII, Pol II0[ser5], l(1)10Ca, Pol II Ser5P, ATSCC2, Pol II[ser2], cohesion-mediated DNA tethering, sister chromatid alignment., Material, H5, CTD, RPII18, RNA Polymerase B, Cistron, delangin, RPII15, DNA-Dependent RNA Polymerase II, dRPB1, RNA PolI 215, dRpb1, DmelCG3284, dRPB9"],"description_synonyms":["sodium salt, Water, l(4)17, l(4)16, l(4)13, Dinitrilotetraacetate, ammonium formate, Gli, Ethylenedinitrilotetraacetic Acid, 13C-labeled, dmMOF, PhrB photolyase activity, YPD, Glukose, Potassium EDTA, H(2)O, A4, Ramp, Myc, magnesium formate, Apaf-1, zinc salt, Solvent, CENP-A, B430311C09Rik, WATER, CENP-C, 1B1, HOH, IDN3, dmTAF[[II]]230, Edetic, Magnesium chloride, Cultural, Grp1, dMOF, B1, 0451/09, 1, 2, 3, myc, sylvite, epithelium, (R*, Disodium Salt, gamma sarcoglycan, SET, 0244/09, Oelsuess, BcDNA:RE21270, hac-1, cel, dm/dMyc, M, N, yeast extract peptone dextrose, ORF19, Saccharomyces uvarum var. melibiosus, BcDNA:AT25108, ecotype, Edetate, SUPPRESSOR OF AUXIN RESISTANCE 3, 2810047L02Rik, DmelCG4299, 671/2, isolation and purification, glucose, salt, Peptidohydrolase Inhibitor, CI, D1, sulfoxide, s, Mycoderma cerevisiae, Gallium EDTA, nickel salt, CiD, Add, Glyceritol, DmCalm, Ce, dTAF[[II]]230, Stars, Ci, Data Set, dmyc1, cobalt (+2) salt, anon-53Fa, Dmel_CG9325, alpha-amino carboxylic acids, Cid, ciD, add, Baker's Yeasts, MgCl2, common salt, sodium (4:1:1) salt, Dm, Cultures, magnesium salt, Algorithm, CG43443, 1316/02, IMD3, BcDNA:HL08040, ci-D, Methionin, Chromium EDTA, Hmet, (alpha-D)-Isomer, dark/hac-1/dapaf-1, D-Glucose, Gro, HTS, Cultural Beliefs, Amino acids, DI-2, CENP-A/CID, CG11628, GLI3FL, CENPA, I-2Dm, Dioxide Snow, formate, NKTL, Baker's Yeast Proteins, S*)-isomer, Pdn, D-Myc, I-2PP1, Applications, Sodium Salt, nitrogen, Taf250, 35kD dystrophin-associated glycoprotein, ammonium (4:1) salt, l(1)16Fg, Edetate Disodium Calcium, Monopotassium chloride, Peptide Peptidohydrolase Inhibitor, STARS, 4-Dimercapto-2, Liquimeth, Ci-155, pds, Ethylenediaminetetraacetic Acid, cohesin localisation to chromatin, CG10798, Gallium, rac-Dithiothreitol, dtl-b, GRP1/cytohesin 1, Endopeptidase Inhibitors, threo-1, Ly62, Cleland's reagent, Pierce, TR-AP, dMYC, Saccharomyces italicus, CG7826, agua, mass-to-charge ratio, CG11633, CenpA, glycyl alcohol, DmelCG17228, Magnesium Disodium, isolation, H2-Qa1, yeast, GP39, Gene Products, CG7835, l(2)k14523, Qa-1(b), Ovhts-RC, MAM, Siah, Technique, lager beer yeast, MAX, anatomical systems, dTAF[[II]]250, FAM39E, T23d, GKLP, Hydrogen chloride, T23b, thallium (+1) salt, cell, Glycerin, rubidium salt, Dmyc, Open Source Software, 2pp2a, Cultural Relativisms, D-CaM, Antagonist, l(3)rJ307, CG10574, S. cerevisiae, Computer Software Application, T20K14_150, Study, dTAF250, Tetracemate, NSC-8400, l(3)05301, sulfoxyl, glycerolum, 2-amino-4-(methylthio)butanoic acid, Protease Antagonists, dSet, Peptide Peptidohydrolase Inhibitors, CG2125, peptidos, SISTER-CHROMATID COHESION 2, 4-(2-hydroxyethyl)-, PTHB1, l(3)j12C8, dark/dapaf-1/hac-1, NTKL, Magnesiumchlorid, 1-(14)C-labeled, Stannous, CG3025, Architectural Barrier, Facility Accesses, Architectural Barriers, copper (+2) salt, Cultural Backgrounds, isosafrole octyl sulfoxide, ACP5, buffer, Digestions, Smp1, Saccaromyces cerevisiae, Dithiothreitol, dTAF230, Kaliumchlorid, Sccharomyces cerevisiae, APAF1, native protein, Temperatures, Proteolyses, 0441/16, carbonyldiamide, Hac-1, C18, cupric formate, Strain, Edathamil, NaCl, aluminum formate, scc2-2, scc2-1, DTL, MET, CG18069, Saccharomyces oviformis, Taf[[II]]250, AU020952, Calcium, DTT, CT16449, sulfox-cide, ensemble, Ethylene Dinitrilotetraacetate, Striated muscle activator of Rho-dependent signaling, Chloride, Magnetic, Monopotassium Salt, azote, ethanenitrile, Magnesium Disodium EDTA, p. pigmentosa retinae, l(3)rK204, plan specification, Computer Programs, Magnesium chloride (MgCl2), Znf198, ten(m), deoxyribonucleate pyrimidine dimer lyase (photosensitive), Peptide Hydrolase Inhibitors, Baker's Yeast, Relativism, CenpA/CID, CCM1, anon-EST:Posey59, CG11452, l(3)rK137, camKII, NCMe, DmelCG6829, classic hairy cell leukaemia, zinc formate, Amino acid, myc-B, DmelCG10798, HEPES Monosodium Salt, KCl, lead salt, Ethylenedinitrilotetraacetic, protein, peptide, cohesin association with chromatin, Tnfsf5, Techniques, DmelCG5723, L-Isomer Methionine, c-MYC, ammonium tetraformate, peptido, Edetic Acid, Klotrix, Ramps, l(4)ar, Apaf1, Hydrolase Inhibitors, protein aggregate, prevention and control, Calcium Tetacine, DmelCG18069, HEPES Monosodium, peptides, TAF-I, l(3)rL201, E927b, dMax, Open, hypoplasia, arc, mAPC, ten[m], ark, l(4)102EFb, DL-glucose, T1, dcaf2, IGAAD, CAMI, lithium salt, DMPROSPER, Tudor repeat associator with PCTAIRE-2, 1-Piperazineethanesulfonic acid, DmelCG10574, CAMC, l(2)k08110, Cenp-A, C2orf34, Facility Access, Tetacine, brewer's yeast, l(2)01103, T5ap, dihydrogen oxide, MGC - 45012, ammonium (2:1) salt, CPVT4, CD40L, Calcium Disodium Edetate, Zfp198, AU016757, D-Apaf-1, l(3)rO534, Pro, [MgCl2], Aminokarbonsaeure, deoxyribonucleic cyclobutane dipyrimidine photolyase activity, CAMKII, 2-Amino-4-(methylthio)butyric acid, Dmel_CG7826, wild leek, StF-IT-1, Th, 4-dimercapto-2, TAFII-250, TAF250/230, Source Softwares, results, Versenate, TAFII250, PROS-1, PROS-2, retinal pigment, Digestion, pro, glycerol, dmax, Computer Applications Softwares, Kaon-Cl 10, cultivar, TNF-related activation protein, WASH, Scc2, nickel (+2) salt, Copper EDTA, Dmel_CG7835, CLNMT, MNB, Yeast, ammonium salt, Distannous, l(2)k06121, Ci/Gli, Step, MS/MS, L2DTL, cobaltous formate, AW743063, CG4299, AW124434, CG17603, TAF[[II]], Methodological Study, acqua, Myc2, R*)-1, Niard, apaf-1, MOF, DMDA, DNA-photoreactivating enzyme, TrATPase, copper salt, IGM, Antiproteases, Qed-1, Wasser, HEPES, 3909, Strains and Sprains, i2pp2a, 1728, CaMK II, SGCG_HUMAN, Dapaf-1/HAC-1, Procedures, TRAP, p50, cesium salt, Aminocarbonsaeure, Brewer's, baker's yeast, dTAFII250, CG9325, CAMIII, Ten[m], alpha-amino acid, Computer, Xt, PHAPII, nipbl, l(3)10419, Potassium Salt, cytohesin/GRP1, dmTAF8, l(4)102ABc, Hac1, Propanetriol, polypeptide chain, dmTAF1, 14C-labeled, p63, p65, HL-VIII, Studies, anon-WO0140519.15, l(2)00634, KMT, Protease Antagonist, l(2)03909, tiny, CD40-L, Pro-Mega, TAF250, CenH3[Cid], PROS, MOS3, l(2)3909, PRECOCIOUS, MS1, MS2, ipp2a2, dapaf-1, CG7128, l(2)SH2 0323, dapaf, calcium formate, Protein Degradations, Physical Barriers, Xylella fastidiosa (strain Temecula1 / ATCC 700964), calcium chloride anhydrous, BHLHE39, X-linked combined immunodeficiency, Camkii, prod, Candida robusta, Chromium Salt, Neural-specific protein-tyrosine phosphatase, TEIF, CG8472, male sterility 1, HIGM1, Controlled., TAF, CG34197, Dicobalt EDTA, Facility, Disodium EDTA, TCEP, C78062, Controlled, small, Applications Software, P105, Open Source, DYRK1, Controlling, TAF[[II]]250, Computer Software, (beta-D)-Isomer, DL-Methionine, protein complex, PCTAIRE2BP, Sprain, igaad, CAMKIIalpha, l(3)84Ab, cenH3, Bp50, Protein Digestion, strontium salt, Magnesium, dCaMKII, group, strain, AI875693, Dithiotreitol, [HCl], calcium/calmodulin-dependent protein kinase II, CAD, GLI3-190, AI461935, Ci155, p230, water, CAM, I2PP2A, Dmel_CG34197, Dyrk1, Dark, Disodium Ethylene, ACETONITRILE, tandem MS, Data Base, CYH1, Accessibility, TAF[[II]]230, PRSS, T20K14.150, Chelaton 3, MALE STERILITY 1 PROTEIN, adducin, Dihydrate, CC1, Tripcellim, CaMK-II, Disodium, dApaf-1/DARK/HAC-1, chromic formate, Protein Gene Products, dSET/TAF-Ibeta, 2610030F17Rik, DmelCG17603, concentration, SCARMD2, AA960152, calcium salt, alpha-amino-gamma-methylmercaptobutyric acid, Qa-1, delangin, Barriers, Chromium, TAF8, EST D, l(3)00844, dAPAF-1, Baker Yeast, TAF1, Qa1, cadmium salt, MeCN, muriate of potash, odd(Oz), Cid[Mel], Ci[D], Physical, calm1, DCAF2, BOUND WATER, MYC, Basodexan, CG17228, oxidane, prevention, RP11-508D10.1, Dicobalt, 1135/09, MYM, 5730420M11Rik, DmelCG9648, 1135/07, anon-EST:Posey9, Polypeptides, protein polypeptide chains, hydrogen chloride, ramp, EG:BACN5I9.1, cobalt(II) formate dihydrate, Method, Protease Inhibitor, CH3-C#N, GRP1, Dextrose, ten-m, Cal49A, RAMP, NUP96, Software Engineering, 5730564G15Rik, 3-Propanetriol, average, ethnicity, type 5 acid phosphatase, amino acids, l(3)rP126, TFIID TAF250, Glycine, DmelCG13176, PTPSTEP, Salt, AI327027, DROPROSA, Saccharomyes cerevisiae, proteins, sulphoxide, CaCl2, T6G21.3, CDLS1, AI047805, set, T-cell antigen Gp39, Wasserstoffchlorid, column, T18c(37), 2-Ethanediylbis(N-(carboxymethyl)glycine), TNFSF5, HtsF, Calcium chloride anhydrous, GPH, Ten-mc, ADD, Physical Barrier, dCaM, Klor-con, Degradations, preventive therapy, CaM KII, R*)-isomer, Tnfrsf5, Dipotassium Salt, CG5723, TAF200, l(2)SH0173, Procedure, collisionally activated dissociation, CaM II, CID, Software Tools, hCD40L, bHLHd7, bHLHd6, bHLHd9, bHLHd8, Computer Applications, H2O, Cultural Background, bHLHd5, bHLHd4, htsRC, Stickstoff, Dm1, Saccharomyces diastaticus, Computer Applications Software, Hydrolase Inhibitor, Chlorwasserstoff, Magnesium Salt, dipyrimidine photolyase (photosensitive), sarcoglycan, 1110049F14Rik, mMyc, CG6829, Hydrogen Oxide, Acid, CAMKIId, Hts, Software Applications, HLA-DR-associated protein II, glycerine, Antagonists, Barrier, CENP-A/Cid, Source Software, Protein Digestions, 1167/13, cromium (+3), Striatum-enriched protein-tyrosine phosphatase, Methodological, N'-1, beta Trypsin, l2dtl, Saccharomyces cerevisiae (Desm.) Meyen ex E.C. Hansen, TAF-IBETA, 2017, TAF-Ibeta, Acids, Endopeptidase, methyl cyanide, chlorane, dMyc1, Disodium Versenate, Carbon Dioxide Snow, Glucose Monohydrate, lead (+2) salt, 0989/01, Brewer's Yeast, TAF230, Computer Software Applications, ADD-87, 5301, 2-amino-4-(methylsulfanyl)butanoic acid, nickel formate dihydrate, Access, dm/myc, lead formate, Glucose, chlorure d'hydrogene, TAPK, Ovhts, aluminum salt, Ethylenediaminetetraacetic, HtsRC, Carbamide, protein-containing complex, thomson, 0763/13, dMyc, Odz, DmelCG3025, CG42273, Nop30, bHLHe57, CaM KMT, Peptide Hydrolase, Prosp, Carmol, gamma-SG, Application, Backgrounds, gp39, CD154, Open Source Softwares, F15E12_6, odz, CATC4, Software Application, Monosodium Salt, beta-Trypsin, DMYc, F23A5.3, uL/min, methanoic acid, Solution, l(3)rH013, PCTAIRE2-binding protein, S cerevisiae, Prodos, max, MGC:45012, 2PP2A, RNCMYC, Nird, Tools, Stannous EDTA, Kochsalz, dSET, Customs, PHKD, bHLHe39, AI256814, Chromatographies, metionina, Anhydrous, dmyc, Potassium, 0671/02, 3-butanediol, DmelCG43443, Hts-RC, Snow, 35 kDa dystrophin-associated glycoprotein, Dry, dApaf-1, Proteins, stepk, Coprin, BG:DS00004.13, chloridohydrogen, potassium formate, alpha-amino acids, acetonitrile, L-Isomer, SPENCDI, Copper, Cell, SGCG, Ten79E, Tool, Versene, polypeptide, l(3)rL433, Wash2, Wash1, l(3)rI160, Natriumchlorid, I-2PP2A, ci[D], Dm I-2, TAF[[II]]250/230, CDLS, Bph, Magnesium chloride anhydrous, ATCMPG1, Met, ATCMPG2, IDN3-B, RPE, lithium formate, DmelCG2125, CenH3, ci155, DMDA1, underdeveloped, prophylaxis, CamKIIalpha, Dark/Dapaf-1/HAC1, Peptidohydrolase Inhibitors, DmelCG11628, ME-IV, Gene Proteins, Peptidase, Applications Softwares, Antiprotease, control, Disodium Ethylene Dinitrilotetraacetate, Relativisms, Attention Deficit Hyperactivity Disorder, TRACP, [OH2], Cultural Relativism, hydrochloric acid, Calcium Salt, Ca2+/calmodulin-dependent protein kinase II, Hac-1/Dark, MGC130048, IPP2A2, Endopeptidase Inhibitor, AU023367, CALML2, DD132, Aminosaeure, Peptide Hydrolase Inhibitor, Monohydrate, Hydrogenchlorid, (DL)-Isomer, pigmented epithelium, Calcium Disodium, Background, c-Myc, halite, Methionine, ARC, Inhibitors, Min, ARK, Computer Program, DmelCG42273, T-BAM, CG13329, c-myc, Edetates, CDPK1, tartrate-resistant acid ATPase, reference sample, scc2, MUB3_18, pigmented retina, min, Computer Programs and Programming, ten-m/odz, Baker, purification, copper, MUB3.18, l(3)j6E2, microlitres per minute, SCAN, Protein Degradation, free, DNA cyclobutane dipyrimidine photolyase activity, Calcitetracemate, Calm, CDT2, cohesin localization to chromatin, Calcium Disodium Versenate, cloruro sodico, preventive measures, reaction, TRAP3, 0320/10, Disodium Calcitetracemate, Methodological Studies, Saccharomyces capensis, Ice, Carbon, Carbon Dioxide, CaM kinase, T18c, Baker's, gamma-sarcoglycan, dihydridooxygen, cdt2, Strains, dApaf1, Racemethionine, Cam1, 37b, 37c, [KCl], phapii, PRE, rock salt, Peptidase Inhibitor, PRO, SG-gamma, Cd40l, aqua, CG9648, classic hairy cell leukemia, Peptide Peptidohydrolase, l(3)05309, CYS, ur, Programs, Program, carbamide, NOP, Distannous EDTA, 3.1.3.48, Softwares, SCC2, and GLY protein 2, Ly-62, retinal pigment layer, cenpA, Mnb, l(1)G0354, Amino Acid, and GLY protein 1, ARABIDOPSIS THALIANA SISTER-CHROMATID COHESION 2, CenH3/CID, Ci/GLI, Voila, Xylella fastidiosa str. Temecula1, l(1)G0359, AI854843, HCl, 0664/07, add-like, hydrogen hydroxide, cdt2-b, common, cdt2-a, hac1, Accesses, d-myc, AI326936, gamma (35kDa dystrophin-associated glycoprotein), natrii chloridum, 2-(methylenedioxy)-4-(2-(octylsulfinyl)propyl)benzene, chloracetamide, dapaf-1S, ATSCC2, phr A photolyase activity, H2NC(O)NH2, Adducin, Mof, Peptidase Inhibitors, dapaf-1L, myc2, D Glucose, STEP, SR3-5, CD40LG, Polypeptide, 2-ethanediylbis(N-(carboxymethyl))-, 7N, MRTL, Inhibitor, gluco-hexose, protein levels, d230, l(1)G0139, Degradation, L Isomer, Gene, Protease, photoreactivating enzyme activity, EfW1, TYPE, Buffer, formic acid, DAGA4, Dark/Hac-1/dApaf1, Karbamid, Glycerine, method, anon-WO03040301.171, potassium salt, Dark/Hac-1/dApaf-1, MYCC, reduced, Ms1, Add-hts, Taf230, method used in an experiment, Trihydroxypropane, Harnstoff, AL024000, 35DAG, c-myc II, L-Methionine, F15E12.6, stratum pigmentosa retinae, Sprains, SCG3, cloruro de hidrogeno, Pedameth, CenH3[CID], CamKII, Taf200, HTS-R1, 0585/13, Ethylene, deoxyribocyclobutadipyrimidine pyrimidine-lyase activity, chlorure de sodium, DmelCG13329, HTS-RC, Taf1p, dark, eau, S cerevisiae Proteins, EMBRYO DEFECTIVE 2773, 5830413P05Rik, lysate, DARK, Trap, taf-ibeta, EDTA, odz/ten-m, piperonyl sulfoxide, N-2-Hydroxyethylpiperazine-N'-2'-ethanesulfonic Acid, liquid, cromium (+3) salt, 1883, culture, dArk, 0563/18, Dapaf-1, F23A5_3, sodium formate, nickel formate, 3H-labeled, Saccharomyces cerevisiae 'var. diastaticus', Glyzerin, CaM, strontium formate, apaf1, Peptide, LGMD2C, l(3)rJ806, Pros, MODIFIER OF SNC1, Software Tool, natural protein, DCK, Protein, Cam, caM, DL-threo-1, deoxyribonucleic photolyase activity, Calcium/calmodulin-dependent protein kinase, Cal, Hydrochloride, TFIID, table salt, 3200001F09Rik, l(2)SH0323, Software, Dark/Apaf-I, cyanomethane, CG13176, DmelCG8472, ZNF198, 3-Trihydroxypropane, Architectural, photolyase activity, uree, cam, Engineering, N 2 Hydroxyethylpiperazine N' 2' ethanesulfonic Acid, TAF[II]250, Amino, Anhydrous Dextrose, nitrogeno, Trypure, H-2T23, DmelCG7128, FIM, [CaCl2], Peptid, SCLL, AA407739, Glc, CID/CENP-A"],"name_synonyms":["cohesin localisation to chromatin, CG10798, determination, AU016757, dm/myc, dmyc1, Saccharomyces cerevisiae 'var. diastaticus', l(1)G0139, Brewer's, myc-B, baker's yeast, DmelCG10798, MYC, Myc, dMyc, dMYC, Saccharomyces italicus, Saccaromyces cerevisiae, B430311C09Rik, nipbl, Baker's Yeasts, Sccharomyces cerevisiae, IDN3, cohesin association with chromatin., anon-WO03040301.171, c-MYC, c-Myc, MYCC, Dm, EG:BACN5I9.1, Saccharomyces diastaticus, chemical analysis, NOP, yeast, c-myc II, ARC, Nop30, bHLHe57, myc, CDLS, mMyc, Scc2, SCC2, scc2-2, lager beer yeast, scc2-1, Saccharomyces oviformis, l(1)G0354, IDN3-B, Yeast, c-myc, ARABIDOPSIS THALIANA SISTER-CHROMATID COHESION 2, T20K14.150, scc2, dm/dMyc, l(1)G0359, Dmyc, DMYc, Saccharomyes cerevisiae, Baker, d-myc, CDLS1, Saccharomyces uvarum var. melibiosus, D-Myc, Myc2, S. cerevisiae, EMBRYO DEFECTIVE 2773, BHLHE39, T20K14_150, cohesin localization to chromatin, Saccharomyces cerevisiae (Desm.) Meyen ex E.C. Hansen, ATSCC2, Niard, S cerevisiae, Candida robusta, RNCMYC, Nird, myc2, Saccharomyces capensis, Baker's, Mycoderma cerevisiae, delangin, assay, 1883, SISTER-CHROMATID COHESION 2, Baker's Yeast, brewer's yeast, dMyc1, bHLHe39, MRTL, Brewer's Yeast, dmyc, Baker Yeast"],"pubmed_abstract_synonyms":["Materials, Chromosome Segregations, l(3)Z23, RNA pol II, 8WG16, Pol-II, Non-Governmental Organizations, RING, protein, RpII215, RpII140[wimp], IDN3, Idn3, Endomycetale, protein polypeptide chains, RNA polII, l(1)L5, Roles, Mediator Like Complex, RNApol2, POL, RNA pol IIo, Concepts, Thyroid Hormone Associated Protein Complex, establishment and maintenance of substrate location, Pol IIo[Ser2], 5, protein aggregate, Cohesin, Chromatins, thymus nucleic acid, Mediator-Like Complex, RNApolII, Man (Taxonomy), SMC complex, scc2, Rpll140, Mediator Transcription Complex, Mediator Like Complexes, ATMED14, proteins, CDLS1, kiaa0892, genetic, Srb-mediator complex, Budding Yeast, Endomycopses, DNA Dependent RNA Polymerase II, Role Concepts, chromosome scaffold, CG3180, polII, CG1554, Double-Stranded DNA, Strains, Ring, CG5595, Segregations, deoxyribonucleic acids, Organizations, DNAn, Gm641, Pol II0[ser2], Budding, nuclear chromatin, Budding Yeasts, Saccharomycetale, establishment and maintenance of substance location, SCC4, RNA Pol II 140, organisation, 4921518A06Rik, aberrant, Modern, Aptitudes, familial, MED14, l(1)DC912, PolII, Double-Stranded, Cohesin Protein Complex, Ability, (Deoxyribonucleotide)n+m, dRing1, RING1, single organism localization, SPBP23A10.01c, Role Concept, Playthings and Play, Dm6, l(3)88Be, Role, Plaything, CSRP, Genetic Materials, scc4, Scc2, Mediator-Like Complexes, DmelCG3180, desoxyribose nucleic acid, SCC2, Genetic Material, pol II, med14, Yeasts, Pol-IIa, DmelCG5595, Yeast, RPB6_DROME, ARABIDOPSIS THALIANA SISTER-CHROMATID COHESION 2, CXorf4, DmelCG1554, EXLM1, Rpb2, Rpb1, RNA Pol II, RPB1, Nongovernmental, F7O18.23, administrative structure, PolIIo, Toys, RNAP II LS, RpABC14, TRAP complex, Pol II0[ser5], Pol II Ser5P, ATSCC2, CRSP150, Chromosome, Material, mau-2, Endomycopsis, H5, ds DNA, Sce/dRing, RPII18, Cistron, inherited genetic, RPII15, DNA, dRPB1, Genetic Material., dRpb1, Strains and Sprains, Endomycetales, humans, DmelCG3284, establishment and maintenance of cellular component location, F7O18_23, dRPB9, MAU2L, atypia, cytoplasmic chromatin, Segregation, Plays, rpII140/wimp, DNS, DRIP150, (Deoxyribonucleotide)n, organization and administration, Gene, CG1163, ENSMUSG00000073278, AU041628, protein-containing complex, dRing, Deoxyribonucleic acids, nipbl, Human, II, dring, Pol II, PolIIc, Deoxyribonucleic Acid, Homo sapiens, polypeptide chain, KIAA0892, Thyroid Hormone-Associated Protein Complex, establishment and maintenance of position, Gene Products, RGR1, dRING, Non-Governmental, atypical, Sprains, Man, Ding, Toy, l(3)RplII140, Organization, Genetic, Playthings, TRAP170, RNAP, Pol II CTD, ligand, Rpll18, RPII215, MEDIATOR COMPONENTS 14, Double Stranded, Deoxyribonucleic acid, dRING1, RNAPII0, BcDNA:RH21608, T16H5_60, 14S cohesin, Ubl, EMBRYO DEFECTIVE 2773, T20K14_150, Abilities, Crsp2, RNAP II, Puppets, chromosome transmission, Non-Governmental Organization, L5, Smc1-Smc3 complex, ARABIDOPSIS MEDIATOR COMPONENTS 14, TRAP (Thyroid Hormone Associated Protein) Complex, CRSP2, l(1)DF912, Play, L mediator complex, RNA Pol II CTD, (Deoxyribonucleotide)m, SISTER-CHROMATID COHESION 2, Pol IIo[ser5], organization, C79399, constitutitional genetic, l(1)G0040, Puppet, RGD1560170, RNA PolII, 9930001L01Rik, Pol IIo, SMC|kleisin ring complex, nuclear cohesin complex, cohesin core heterodimer, Dring, organizational structure, protein complex, DNAn+1, Pol IIc, DRING, Proteins, Sprain, Pol IIa, TR-Associated Protein (TRAP) Complex, STRUWWELPETER, defective, Cistrons, RNAPII, Concept, DmelCG1163, RPII140, RPB2_DROME, native protein, natural protein, CG3284, T16H5.60, Protein, CDK8-containing TRAP/mediator complex, Strain, RNA Pol IIc, SMC/kleisin ring complex, Ring/Sce, establishment and maintenance of localization, DmRP140, ds-DNA, CDLS, sister chromatid alignment, Nongovernmental Organizations, Rpll215, scc2-2, scc2-1, administrative management, IDN3-B, wimp, Trap170, RP140, T20K14.150, Rnf2, Pol IIo[ser2], Talents, single-organism localization, RpII, l(1)10Ca, Protein Gene Products, 9S cohesin, Gene Proteins, cohesion-mediated DNA tethering, Pol II[ser2], Non Governmental Organizations, localisation, ORF1, Talent, Modern Man, Very poor growth, Desoxyribonukleinsaeure, 4933421G18Rik, organizational management, CTD, RNA Polymerase B, SCE, delangin, DNA-Dependent RNA Polymerase II, RNA PolI 215, Nongovernmental Organization, hereditary"],"citation_count":["0"],"additional_accession":["PXD022100"]},"is_claimable":false,"name":"LC/MS Analysis of Myc-Affinity Captured, on-Beads Crosslinked, and on-Beads Digested S. cerevisiae SCC2 subunit of cohesin loading factor","description":"Myc-Affinity Capture and on-Beads Crosslinking\nFor each Saccharomyces cerevisiae strain (C-terminally myc-tagged Scc2 subunit of the cohesin loading factor complex or wild-type BY4741), 2x 3 L YPD medium (add 300 ml 20% Glucose to each 3 L of YPD before using) were separately inoculated in a 5L flask with 15 ml overnight culture and incubated at 30C with shaking until OD600 reached 1.5. Cell pellets were resuspended in up to 30 ml in BH0.15 extraction buffer (25 mM HEPES, pH 7.5, 2 mM MgCl2, 0.1 mM EDTA, 0.5 mM EGTA-KOH, 15% Glycerol, 0.1% NP-40, 150 mM KCl) with freshly added 100x protease inhibitor and 5mM DTT. Cells were lysed using liquid nitrogen and dry ice. \nPierce Anti-c-Myc Magnetic Beads (150 ul) were washed with 1 ml of BH0.15 extraction buffer. The supernatant was discarded, and the washed beads were added to the protein lysate and incubated on a rotating wheel overnight at 4C. Proteins bound to anti-Myc beads were washed twice with 15 ml BH0.15. Beads were washed with 1 ml of pre-elution rinse buffer (50 mM HEPES, pH 7.5, 75 mM KCl, 1 mM EGTA). Proteins bound to anti-myc beads were crosslinked by adding 150 ul pre-elution rinse buffer complemented with protease inhibitor cocktail and DTT with 0.6 ul of 250 mM disuccinimidyl sulfoxide (DSSO) to a final concentration of 1mM and let to crosslink at room temperature for 40 min. The crosslinking reaction was quenched by adding 7.5 ul of 1 M NH4CO3 (final 50 mM) and rotating at RT for 15 min. \n\nOn-Beads Protein Digestion\nCrosslinked proteins bound to anti-myc beads were washed twice with 500 ul 10 mM Tris-HCl pH7.5, 150 mM NaCl, then denatured, reduced and digested with 100 ul 50 mM Tris-HCl pH7.5, 2 M urea, 1 mM TCEP (Pierce), 5 ul Trypsin at 0.1 ug/ ul (Sequencing Grade Modified Trypsin; Promega), at 30 C for 30 min. The supernatant was collected. Another 60 ul of 50 mM Tris-HCl pH7.5, 2 M urea, 5 mM 2-Chloroacetamide (CAM, Sigma) were added to the beads to alkylate free cysteines, and the resulting supernatant combined to the first one.  CAM was next added to 2.5 mM and the reaction incubated in the dark at RT for 30 min. CaCl2 was added to 2 mM along with 5 ul of Trypsin, and the digestion was let to proceed at 37C overnight. The digestion was quenched by adding formic acid to 5%. The combined supernatants from these multiple steps constitute the first elution (E1). These digestion steps were repeated once, and the collected supernatants combined as the second elution (E2). \n\nLC/MS Acquisition\nDigested peptides were analyzed on an Orbitrap Fusion Lumos mass spectrometer equipped with a FAIMS Pro interface coupled to a Dionex Ultimate 3000 RSCLnano System.  Peptides were loaded (5 or 10ul for the Scc2-myc digests and 5 or 66ul for the wild-type BY4741 negative control digests) on an Acclaim PepMap 100 C18 0.3 mm i.D. x 5 mm length trap cartridge with loading pump at 2 ul/min via autosampler.  A 75 um i.d. analytical microcapillary column was packed in-house with 250 mm of 1.9 um ReproSil-Pur C18-AQ resin. Column temperature was maintained at 40C.  The organic solvent solutions were water/acetonitrile/formic acid at 95:5:0.1 (v/v/v) for buffer A (pH 2.6) and at 20:80:0.1 (v/v/v) for buffer B.  The chromatography gradient was a 25 min column equilibration step in 2% B; a 3 min ramp to reach 10% B; 90 min from 10 to 40 % B; 6 min to reach 95% B; a 9 min wash at 95% B; 0.1 min to 2% B; followed by a 12 min column re-equilibration step in 2% B.  The nano pump flow rate was set to 180 nL/min.  Orbitrap Fusion Lumos was set up with peptide identification method as: full MS1 resolution 120,000; ITMS2 isolation window 1.4 m/z, ITMS2 max ion injection time 50 ms, ITMS2 CID 35% with normal scan.  FAIMS compensation voltages (CVs) were set up as -40V, -60V, and -80V.  \n\nMS Dataset Processing\nCollected MS/MS spectra were searched with the ProLuCID algorithm against a database of 12276 protein sequences combining 6010 non-redundant Saccharomyces cerevisiae proteins (NCBI, 2017-05-16 release), 193 common contaminants, and their corresponding 6138 randomized amino acid sequences.  All cysteines were considered as fully carboxamidomethylated (+57 Da statically added), while methionine oxidation was searched as a differential modification.  DTASelect v1.9 and swallow, an in-house developed software, were used to filter ProLuCID search results at given FDRs at the spectrum, peptide, and protein levels.  Here, all controlled FDRs were less than 1%.  ","dates":{"publication":"Tue Oct 20 17:29:00 BST 2020"},"accession":"MSV000086339","cross_references":{"pubmed":["35654035"]}}