Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/2019MQ005_SUBO_30_3_1ug.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/2019MQ005_SUBO_37_2_1ug.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/2019MQ005_SUBO_37_1_1ug.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/2019MQ005_SUBO_42_1_1ug.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/2019MQ005_SUBO_37_3_1ug.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/2019MQ005_SUBO_42_2_1ug.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/2019MQ005_SUBO_42_3_1ug.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/2019MQ005_SUBO_30_1_1ug.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/2019MQ005_SUBO_30_2_1ug.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/sp_spombe_2019_06.fastaftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/02/PXD014400/2019MQ005_SUBO_001to009_XLtest.pdResultprimary200OKelena.hidalgo@upf.eduSusanna BoronatMass SpectrometryShotgun proteomicsLc-msmsAggregatesS. pombeInsoluble proteinHeat stresshttps://www.ebi.ac.uk/pride/archive/projects/PXD014400Biological triplicates of pellet fractions of 250 ml cultures (OD600 0.5) grown in MM at 30°C, 37°C or 42°C were obtained using a Precellys cell disrupter (Bertin Technologies) to lyse the cell suspensions. 10 µg of each pellet fraction were reduced with dithiothreitol (100 mM, 37ºC, 60 min) and alkylated in the dark with iodoacetamide (5 µmol, 25ºC, 20 min). The resulting protein extract was washed with 2 M urea in 100 mM Tris-HCl and then with 50 mM ammonium bicarbonate for digestion with endoproteinase LysC (1:10 w:w, 37ºC, o/n) and then for trypsin digestion (1:10 w:w, 37ºC, 8h, Promega). After digestion, peptide mix was acidified with formic acid and desalted with a MicroSpin C18 column (The Nest Group, Inc) prior to LC-MS/MS analysis. Samples were analyzed using a LTQ-Orbitrap XL mass spectrometer (Thermo Fisher Scientific, San Jose, CA, USA) coupled to an EASY-nLC (Thermo Fisher Scientific (Proxeon), Odense, Denmark). Peptides were loaded onto the 2-cm Nano Trap column with an inner diameter of 100 μm packed with C18 particles of 5 μm particle size (Thermo Fisher Scientific) and were separated by reversed-phase chromatography using a 12-cm column with an inner diameter of 75 μm, packed with 3 μm C18 particles (Nikkyo Technos Co., Ltd. Japan). Chromatographic gradients started at 97% buffer A and 3% buffer B with a flow rate of 300 nl/min for 4 minutes and gradually increased to 5% buffer B and 95% buffer A in 1 min and to 65% buffer A and 35% buffer B in 120 min. After each analysis, the column was washed for 10 min with 10% buffer A (0.1% formic acid in water) and 90% buffer B (0.1% formic acid in acetonitrile). The mass spectrometer was operated in positive ionization mode with nanospray voltage set at 2 kV and source temperature at 200°C. The acquisition was performed in data-dependent acquisition (DDA) mode and full MS scans with 1 micro scans at resolution of 60,000 were used over a mass range of m/z 350-1500 with detection in the Orbitrap mass analyzer. Auto gain control (AGC) was set to 1E5, dynamic exclusion (90 seconds) and charge state filtering disqualifying singly charged peptides was activated. In each cycle of DDA analysis, following each survey scan, the top ten most intense ions with multiple charged ions above a threshold ion count of 5,000 were selected for fragmentation. Fragment ion spectra were produced via collision-induced dissociation (CID) at normalized collision energy of 35% and they were acquired in the ion trap mass analyzer. AGC was set to 1E4, isolation window of 2.0 m/z, activation time of 30 ms and maximum injection time of 100 ms was used. All data were acquired with Xcalibur software v2.1.PrideAcquired spectra were analyzed using the Proteome Discoverer software suite (v2.0, Thermo Fisher Scientific) and the Mascot search engine (v2.6, Matrix Science). The data were searched against a Swiss-Prot Schizosaccharomyces pombe reference proteome database (as in June 2019) plus a list of common contaminants and all the corresponding decoy entries (Beer et al, 2017). For peptide identification a precursor ion mass tolerance of 7 ppm was used for MS1 level, trypsin was chosen as enzyme and up to three missed cleavages were allowed. The fragment ion mass tolerance was set to 0.5 Da for MS2 spectra. Oxidation of methionine and N-terminal protein acetylation were used as variable modifications whereas carbamidomethylation on cysteines was set as a fixed modification. Peptide quantification data were retrieved from the “Precursor ion area detector” node from Proteome Discoverer (v2.0) using 2 ppm mass tolerance for the peptide extracted ion current (XIC). The obtained values were used to calculate protein fold-changes and their corresponding adjusted p- and q-valuesProteomicsElena HidalgoOxidative Stress and Cell Cycle Group, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, 08003 Barcelona, SpainPARTIALSchizosaccharomyces Pombe 927susanna.boronat@upf.edu32075773 Cabrera M, Boronat S, Marte L, Vega M, Pérez P, Ayté J, Hidalgo E. Chaperone-Facilitated Aggregation of Thermo-Sensitive Proteins Shields Them from Degradation during Heat Stress. Cell Rep. 2020 30(7):2430-2443.e4 10.1016/j.celrep.2020.01.077Universitat Pompeu FabraSpainCells have developed protein quality-control strategies to manage the accumulation of misfolded substrates during heat stress. Using a soluble reporter of misfolding in fission yeast, Rho1.C17R-GFP, we demonstrate that upon mild heat shock, the reporter collapses in protein aggregate centers (PACs). They contain and/or require several chaperones, such as Hsp104, Hsp16, and the Hsp40/70 couple Mas5/Ssa2. Stress granules do not assemble at mild temperatures and, therefore, are not required for PAC formation; on the contrary, PACs may serve as nucleation centers for the assembly of stress granules. In contrast to the general belief, the dominant fate of these PACs is not degradation, and the aggregated reporter can be disassembled by chaperones and recovers native structure and activity. Using mass spectrometry, we show that thermo-unstable endogenous proteins form PACs as well. In conclusion, formation of PACs during heat shock is a chaperone-mediated adaptation strategy.Chaperone-Facilitated Aggregation of Thermo-Sensitive Proteins Shields Them from Degradation during Heat Stress.Cabrera Margarita M, Boronat Susanna S, Marte Luis L, Vega Montserrat M, Pérez Pilar P, Ayté José J, Hidalgo Elena Esodium salt, Water, l(4)17, l(4)13, ammonium formate, Gli, 13C-labeled, Size, cadmium salt, MeCN, ion, Cid[Mel], fond, Ci[D], Particle, H(2)O, BOUND WATER, Basodexan, magnesium formate, CycEI, Apaf-1, oxidane, zinc salt, CENP-A, prevention, Long Term, WATER, CENP-C, HOH, 5730420M11Rik, dmTAF[[II]]230, Polypeptides, hydrogen chloride, cobalt(II) formate dihydrate, CH3-C#N, B1, 1, 2, Software Engineering, Faroe Islands, WMS, NICR2359, (R*, SET, 2810441M03Rik, Divorced, BcDNA:RE21270, TFIID TAF250, hac-1, cel, Divorces, AI047805, DmelCG4299, isolation and purification, set, Wasserstoffchlorid, CI, column, D1, n, z, DmAAF34715, nickel salt, SGS, CiD, LC-MS2, 1E4, Ce, dTAF[[II]]230, preventive therapy, Ci, Bonin Islands, cobalt (+2) salt, anon-53Fa, TAF200, Longterm Effect, l(2)SH0173, Cyc E, not genetically inherited, collisionally activated dissociation, CID, Cid, ciD, Software Tools, ACMICD, Computer Applications, H2O, sodium (4:1:1) salt, BG:DS07108.3, count, Dm1, magnesium salt, Computer Applications Software, Chlorwasserstoff, Sizes, ci-D, l(2)05206, CG6829, activation, Hydrogen Oxide, dark/hac-1/dapaf-1, Software Applications, HLA-DR-associated protein II, DI-2, CENP-A/Cid, CENP-A/CID, CENPA, Source Software, I-2Dm, formate, cromium (+3), beta Trypsin, I-2PP1, Applications, TAF-IBETA, Taf250, liquid chromatography-tandem mass spectroscopy, ammonium (4:1) salt, TAF-Ibeta, methyl cyanide, chlorane, lead (+2) salt, 4-Dimercapto-2, Ci-155, TAF230, Cleland Reagent, Particle Sizes, rac-Dithiothreitol, nickel formate dihydrate, cycline, lead formate, Effects, threo-1, chlorure d'hydrogene, Cleland's reagent, DmcyclinE, aluminum salt, 4-dimercapto-, Carbamide, R*)-, NAT13P, thomson, CG7826, agua, mass-to-charge ratio, cdi7, CenpA, cyclinE, isolation, IT, LC-MSMS, CG7835, Cdi7, CDI7, CG42273, Carmol, nat13, Siah, Application, Open Source Softwares, dTAF[[II]]250, Hydrogen chloride, thallium (+1) salt, Longterm, cell, Software Application, beta-Trypsin, rubidium salt, Open Source Software, 2pp2a, Long-Term, methanoic acid, DmcycE, ions, CG10574, Computer Software Application, Injectable, dTAF250, Faeroe Islands, 2PP2A, Tools, dSET, dSet, CG2125, peptidos, Chromatographies, PTHB1, 3-butanediol, dark/dapaf-1/hac-1, grupo, 1-(14)C-labeled, dApaf-1, mak3, Clelands Reagent, BG:DS00004.13, chloridohydrogen, potassium formate, copper (+2) salt, backward, acetonitrile, buffer, Cell, Dithiothreitol, Tool, dTAF230, polypeptide, APAF1, l(2)k02514, DmCycE, Hac-1, carbonyldiamide, I-2PP2A, ci[D], C18, cupric formate, chemical analysis, Dm I-2, Long Term Effects, TAF[[II]]250/230, MFS1, MIXL, aluminum formate, DTL, Cleland's, Taf[[II]]250, AU020952, DTT, lithium formate, l(2)k02602, DmelCG2125, CenH3, ci155, underdeveloped, ensemble, increased number, prophylaxis, Separations, Dark/Dapaf-1/HAC1, ethanenitrile, Longterm Effects, ME-IV, Cleland's Reagent, Computer Programs, D-CycE, Applications Softwares, 1E4 细胞, control, liquid chromatography tandem mass spectrometry, Mak3, [OH2], CenpA/CID, hydrochloric acid, reversed, MAK3, liquid chromatography tandem mass spectroscopy, Hac-1/Dark, IPP2A2, NCMe, determination, DmelCG6829, classic hairy cell leukaemia, zinc formate, atado, lead salt, Hydrogenchlorid, peptide, ammonium tetraformate, peptido, Ccne, Min, Apaf1, ARK, prevention and control, Effect, Injectables, Computer Program, DmelCG42273, MIX, CG13329, umol, increased, LCMSMS, peptides, reference sample, TAF-I, E927b, Open, hypoplasia, min, arc, Computer Programs and Programming, mAPC, analyzer, purification, CG12352, copper, ark, SCAN, T1, hNAT5, preventive measures, IGAAD, µmol, lithium salt, hSAN, Sputolysin, DmelCG10574, Cenp-A, dihydridooxygen, dApaf1, dihydrogen oxide, GPHYSD2, ammonium (2:1) salt, Long-Term Effects, Mak3p, phapii, D-Apaf-1, NAT13, LC-MS/MS, aqua, Dmel_CG7826, StF-IT-1, Th, 4-dimercapto-2, TAFII-250, classic hairy cell leukemia, TAF250/230, br37, Nat13, ur, Source Softwares, Programs, Program, carbamide, TAFII250, Cleland, Ionen, Computer Applications Softwares, Softwares, nickel (+2) salt, Dmel_CG7835, cenpA, Mnb, MNB, MILD1, ammonium salt, CenH3/CID, Ci/Gli, Ci/GLI, HCl, cobaltous formate, hydrogen hydroxide, 3-Butanediol, hac1, CG4299, MASS, AW124434, CG17603, TAF[[II]], acqua, dapaf-1S, R*)-1, nanospray, H2NC(O)NH2, apaf-1, dapaf-1L, l35Dd, SR3-5, Acetamide, Computer Software Applications., copper salt, Polypeptide, Wasser, i2pp2a, 1728, 2600005K24Rik, accessory, Gruppe, d230, nat5, Dapaf-1/HAC-1, Reagent, cesium salt, Z-350, FBN, dTAFII250, NAT5P, cycE, Computer, EfW1, supernumerary, l(2)br37, PHAPII, LC-MS-MS, CYCLE, formic acid, Buffer, 4-(3-(4-(1-(4-methylphenyl)-3-(4-(2-methoxyphenyl)piperazine-1-yl)propoxy)benzoyl)indole-1-yl)butyric acid, Dark/Hac-1/dApaf1, Karbamid, l(4)102ABc, Hac1, potassium salt, Dark/Hac-1/dApaf-1, reduced, dmTAF1, Taf230, ECTOL1, Injection, 14C-labeled, Harnstoff, tiny, Separated, cloruro de hidrogeno, Pro-Mega, TAF250, CenH3[Cid], CenH3[CID], Taf200, CYCE, DmelCG3938, iones, CyclE, ipp2a2, dapaf-1, 3938, DmelCG13329, Taf1p, dapaf, dark, eau, calcium formate, OCTD, l(2)k05007, X-linked combined immunodeficiency, dm-cycE, 10^[-6], Suspension, DARK, grupos, taf-ibeta, DmelCG12352, NAT5, Nat5, cromium (+3) salt, Long-Term Effect, TAF, dArk, Dapaf-1, 2-iodo-, sodium formate, Controlled, span, small, Applications Software, Open Source, DYRK1, Controlling, TAF[[II]]250, Computer Software, nickel formate, 3H-labeled, igaad, strontium formate, l(3)84Ab, cenH3, apaf1, strontium salt, Peptide, group, Dithiotreitol, [HCl], AW112078, LC/MS/MS, Software Tool, CAD, Ci155, p230, water, DL-threo-1, I2PP2A, Hydrochloride, TFIID, Dyrk1, Dark, CyeE, Software, Dark/Apaf-I, ACETONITRILE, cyanomethane, WMS2, SAN, San, Separation, 10^[-9], TAF[[II]]230, PRSS, uree, CC1, l(2)35Dd, Engineering, Tripcellim, Rest, TAF[II]250, 1E4 cell, dApaf-1/DARK/HAC-1, chromic formate, san, Trypure, present in greater numbers in organism, dSET/TAF-Ibeta, 2610030F17Rik, DmelCG17603, Ion, SSKS, calcium salt, Peptid, assay, AA407739, CG3938, groupe, dAPAF-1, TAF1, CID/CENP-Aliquid chromatography tandem mass spectroscopy, LC-MS2, Temperature, LC/MS/MS, LCMSMS, Temperatures, liquid chromatography-tandem mass spectroscopy, Hot, LC-MSMS, particle-bound, LC-MS/MS, Heat, Hot Temperatures, liquid chromatography tandem mass spectrometry, LC-MS-MS.IPP2A2, ion, Ass-1, Fission, protein, Schizosaccharomyces pombe, 5730420M11Rik, peptide, Polypeptides, L-Isomer Methionine, Fission Yeast, Drug Tolerance, Methionine, peptido, dorsal marginal zone, fold, Software Engineering, protein aggregate, fission yeast, Computer Program, WMS, C79691, SET, peptides, TAF-I, M, Open, E430039A18Rik, Computer Programs and Programming, T6G21.3, Lccp, DmelCG4299, ASS, IGAAD, set, DmelCG10574, ppm, GPHYSD2, Racemethionine, drug tolerance, SGS, phapii, Stars, immune system tolerance, T5E21.12, 2-Amino-4-(methylthio)butyric acid, StF-IT-1, Search, S pombe, not genetically inherited, Source Softwares, Software Tools, Programs, ACMICD, Program, Computer Applications, Ionen, Computer Applications Software, Computer Applications Softwares, Immunologic Tolerance, Henson's node, Methionin, Softwares, Hmet, T5E21_12, parent ion, Yeast, Software Applications, HLA-DR-associated protein II, DI-2, MS/MS, Source Software, I-2Dm, common, MASCOT, MASS, CG4299, beta Trypsin, I-2PP1, Applications, Self Tolerance, TAF-IBETA, precursor ion, Tolerance, Biocatalyst, DMZ, TAF-Ibeta, Polypeptide, STARS, i2pp2a, Liquimeth, Proteomes, Computer Software Applications, 2-amino-4-(methylsulfanyl)butanoic acid, AW549739, Biocatalysts, L Isomer, FBN, Gene, 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Spectrum Analysis, Dmrho, SVE, single-organism biosynthetic process, Heat-Stress Reaction, reference sample, Analyses, FATE, catabolism, DMRHO, DmelCG10578, Ssa2, SSA2, rhoA, RHO1, Rho1, preventive measures, DmelCG8416, PACs, rho1, biotransformation, PAC, atrial ectopic beats, Aggregate, A530054J02Rik, PAS, preventive therapy, Heat Shock Response, anon-WO0172774.135, DmRHO-A, Rho-1, Crosti-gianotti syndrome, RHOb, CG1004, Heat-Shock Responses, infections, S pombe, Spectrum Analysis, Heat Stresses, Dm Rho1, RORNP, rhom, Spectroscopy, Cultural Background, RhoN19, Heat Shock Stresses, soluble, Cultures, Ectopic supraventricular rhythms, RhoA, ve, CT43, Protein Aggregate, secretion, Yeast, 1810007I17Rik, Cultural Beliefs, D-Rho1, Ve, Heat-Stress Reactions, CG8416, Spectrometry, dhdj1, RHO, Rho, anon-WO0140519.166, Granule, DMRHOa, DMRHOb, hDj-1, rhomboid/veinlet, acrodermatitis, Heat Stress Response., MAS5, rho, dHDJ1/HSP40, Hdj1, HDJ1, Dmelrho, DRORHO, conformation, Gene, Spectrum Analyses, Schizosaccharomyces malidevorans, Atrial premature complex, Heat shock 40 kDa protein 1, protein-containing complex, Rho GTPase, Gianotti Crosti syndrome, cellular catabolism, Human DnaJ protein 1, YG102, DnaJ1 64EF, papular infantile, DRhoA, Gene Products, Mass, Sis1, Neisseria gonorrhoeae, Mass Spectroscopy, Heat Stress Response, Backgrounds, CG10578, Schizosaccharomyces pombeP, dRhoA, breakdown of chemical, Cultural Relativisms, DRho1, Atrial ectopic beats, DrhoA, ectopic supraventricular rhythm, DNAJ-1, 0610007I11Rik, Heat Shock Reaction, Drho1, Customs, dRho1, Heat shock protein 40, Controlled, cellular breakdown, Controlling, Rho kinase, Stress Granule, AI646302, degradation, protein complex, DROJ1, l(2)52Fa, Proteins, iks, RO60, l(2)k02107b, Cultural Backgrounds, infantile lichenoid, DroJ1, Cell, supraventricular ectopy, dhdj-1, SS-A|Ro, MS, Heat Stress, native protein, Temperatures, Protein, Heat Stress Reaction, dnRho, DNAJ1, Mass Spectrum Analyses, breakdown of molecule, Hsp-40, Mass Spectrum, Heat-Shock Reactions, biodegradation, prophylaxis, Hsp40, RSPH16B, E3.10/J3.8, Premature supraventricular beats, Heat Shock, Heat Shocks, droj1, Protein Gene Products, Ssa, Gene Proteins, breakdown of substance, Fission Yeasts, Heat-Stress Responses, control, Stress, hsp40, Relativisms, Relativism, Cultural Relativism, n(2)k07236, Heat-Stress Response, General activity, DnaJ protein homolog 1, HSP40biochemical pathways, Shock, cellular breakdown, Heat Shock Stress, Heat-Shock Reaction, Heat Shock Response, Heat-Stress Reaction, Heat-Shock Reactions, biodegradation, degradation, breakdown of chemical, catabolism, Proteins, Heat-Stress Reactions, Gene, Heat-Shock Responses, Heat Shock, Heat Shocks, cellular catabolism, Heat Stresses, Protein Gene Products, allergic reaction, Gene Proteins, breakdown of substance, Heat-Stress Responses, Heat Stress, Heat Shock Stresses, sensitive, Heat Shock Reaction, Protein, Stress, Gene Products, cellular degradation, biotransformation, Heat, Heat Stress Reaction, secretion, Heat Stress Response., sensitivity, Heat-Stress Response, breakdown of moleculefalseInsoluble fraction_Heat stress_S.pombe LC-MSMSWe have carried out proteomic studies to identify the thermo-unstable proteome fraction in fission yeast. We have performed biochemical separation of pellet fractions by centrifugation of whole cell extracts from wild-type cultures grown at different temperatures, and performed untagged liquid chromatography coupled to tandem MS (LC-MS/MS) of biological triplicates. We demonstrate that formation of protein aggregates during heat shock is a chaperone-mediated adaptation strategy which occurs with thermo-unstable endogenous proteins as well.2020-02-242019-06-26PXD014400NEWT:6945NEWT:3555NEWT:241368NEWT:2NEWT:157546NEWT:190802NEWT:35554NEWT:9778NEWT:150475NEWT:9417NEWT:347515NEWT:1216979NEWT:307972NEWT:32046NEWT:544496NEWT:5180NEWT:256737NEWT:2042546NEWT:115104NEWT:1081927NEWT:67825NEWT:43179NEWT:13076NEWT:1249668NEWT:376741NEWT:317NEWT:1736309NEWT:7227NEWT:7469NEWT:885318NEWT:415540NEWT:876138NEWT:4081NEWT:554NEWT:98334NEWT:237561NEWT:6928NEWT:10036NEWT:7574NEWT:1351NEWT:7215NEWT:272563NEWT:507601NCBITaxon:79857NCBITaxon:6157NEWT:95648NEWT:3885NEWT:746360NEWT:6239NEWT:1589NEWT:470150NEWT:135622NEWT:216257NEWT:6915NEWT:9986NEWT:101510NEWT:4054NEWT:3880NEWT:3641NEWT:8782NEWT:1000589NEWT:1902NEWT:85962NEWT:160488NEWT:28104NEWT:317447NEWT:7955NCBITaxon:2NEWT:985076NEWT:7959NEWT:2261NEWT:4565NEWT:1264690NEWT:6192NEWT:28532NCBITaxon:38727NEWT:34305NEWT:59729NCBITaxon:183674NEWT:224308NEWT:626528NEWT:139927NEWT:4558NEWT:209285NEWT:216595NEWT:243230NEWT:8355NEWT:1283NEWT:931281NEWT:7029NEWT:1283300NEWT:6183NEWT:334747NEWT:61235NCBITaxon:79824NEWT:4787NCBITaxon:4563NEWT:5755NEWT:3218NEWT:5759NEWT:1736231NEWT:436486NEWT:6287NEWT:2242NEWT:300641NEWT:4784NEWT:727NEWT:9796NEWT:725NEWT:360106NEWT:260707NEWT:287NEWT:10117NEWT:10239NCBITaxon:6191NEWT:10116NEWT:1280NEWT:1836NEWT:1735272NEWT:83334NEWT:185431NEWT:83332NEWT:29760NEWT:260704NEWT:703612NEWT:260705NEWT:80863NEWT:44685NEWT:2697049NEWT:1148NEWT:11676NEWT:55571NEWT:100226NCBITaxon:6073NEWT:4530NEWT:4896NEWT:6279NEWT:1123869NEWT:7370NEWT:75058NEWT:83906NEWT:607699NEWT:6282NEWT:208964NEWT:1134506NEWT:575584NEWT:1773NEWT:38783NEWT:8727NEWT:1895NEWT:1182590NEWT:8726NEWT:10090NEWT:935293NEWT:64152NEWT:749200NEWT:4120NEWT:51515NEWT:5693NEWT:8724NEWT:51511NEWT:92867NEWT:8723NEWT:990346NEWT:5334NEWT:145953NEWT:257309NEWT:230741NEWT:284812NCBITaxon:10359NCBITaxon:1313NEWT:43330NEWT:242619NEWT:44544NEWT:632957NEWT:373995NEWT:5689NEWT:544404NEWT:3702NEWT:129249NEWT:9925NEWT:8839NEWT:4232NEWT:990119NEWT:2758385NEWT:4113NEWT:837NEWT:11298NEWT:171101NEWT:196627NEWT:408172NEWT:5691NEWT:408170NEWT:493760NEWT:260710NEWT:627025NEWT:400772NEWT:1097677NEWT:3708NEWT:128161NEWT:106592NEWT:1117957NEWT:9913NEWT:1432138NEWT:10312NEWT:1424507NEWT:4100NEWT:1076NEWT:6763NEWT:803NEWT:8030NEWT:29722NEWT:380394NEWT:1692259NEWT:1639NEWT:188229NEWT:3818NEWT:480NEWT:4909NEWT:180066NEWT:67767NEWT:46835NEWT:135588NEWT:1843183NEWT:95486NEWT:58002NEWT:9103NEWT:4577NEWT:5664NEWT:2157NEWT:146479NEWT:10306NEWT:1911079NEWT:8022NEWT:145943NEWT:3635NEWT:5811NEWT:235443NEWT:1480154NEWT:1274414NEWT:59202NEWT:3197NEWT:9615NEWT:10299NEWT:860688NEWT:884019NEWT:169963NEWT:36329NEWT:1147787NEWT:72407NEWT:9606NEWT:367830NEWT:157295NEWT:178616NEWT:410289NEWT:373153NEWT:915099NEWT:74940NEWT:1450511NEWT:360094NEWT:470NEWT:84023NEWT:9838NCBITaxon:9615NEWT:58334NEWT:1193501NEWT:3055NEWT:6326NEWT:6689NEWT:2762NEWT:5476NEWT:1174673NEWT:562NEWT:33952NEWT:1274432NEWT:1274426NEWT:1423NEWT:4932NEWT:70448NEWT:9825NEWT:1274423NEWT:3603NEWT:698936NEWT:2759NEWT:3847NEWT:39946NEWT:9823NEWT:9940NEWT:327160NEWT:573NEWT:9031NEWT:1274420NEWT:7091320757730000-0003-1745-1556