Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_1WT_20_2h_n4_1_1212.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_1WT_20_2h_n5_2_1225.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_1WT_20_2h_n3_1_1200.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_3D8_20_2h_n3_3_1208.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_3D8_20_2h_n4_2_1219.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_3D8_20_2h_n5_2_1231.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_1WT_20_2h_n5_1_1224.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_3D8_20_2h_n3_2_1207.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_3D8_20_2h_n5_1_1230.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_1WT_20_2h_n4_3_1214.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_3D8_20_2h_n4_1_1218.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_1WT_20_2h_n5_3_1226.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_3D8_20_2h_n3_1_1206.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_1WT_20_2h_n3_3_1202.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_1WT_20_2h_n4_2_1213.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_3D8_20_2h_n4_3_1220.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_1WT_20_2h_n3_2_1201.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/20141119_AlCH_3D8_20_2h_n5_3_1232.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/MaxQuantOutput.tar.gzftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/01/PXD001584/andromeda_APL.zipprimaryOK200003320chiara.guerrera@inserm.frChiara guerreraMass SpectrometryShotgun proteomicsTularemiaLabelfreeF. tularensisQ exactive plusArginine transporterhttps://www.ebi.ac.uk/pride/archive/projects/PXD001584MS/MS Protein Identification and Quantification WT and ∆argP mutant strains were grown overnight in CDMmin. The next day, bacteria were resuspended at an OD600 of 0.1 in CDMmin or in CDM containing arginine at the concentration of 0.43 mM and cultivated for 1 h or 2 h. The cells (50 mL) were harvested by centrifugation at 3,000 x g for 20 min and resuspended in 2 mL of 5 mM Tris-HCl pH 8. The bacterial suspensions were sonicated three times for 30 s at 4.0 output, 70% pulsed (Branson Sonifier 250). Lysates were centrifuged once again at 8,000 x g for 6 min and supernatant were collected. For label-free, relative quantitative analysis, 50 µg protein from each of the eight samples (WT and ∆argP, at 1 h and 2 h of growth, at 2.3 mM or 0.46 mM arginine) were precipitated in 80%, acetone and digested with 1 µg sequencing grade trypsin (Promega). Following an overnight digestion at 37°C samples were vacuum dried, and resuspended in 100 µL of 10% acetonitrile, 0.1% formic acid, for LC-MS/MS. For each run, 5 µL were injected in a nanoRSLC-Q Exactive PLUS (Dionex RSLC Ultimate 3000, Thermo Scientific, Waltham, MA, USA). Peptides were separated on a reversed-phase liquid chromatographic column (Pepmap C18, Dionex). Chromatography solvents were (A) 0.1% formic acid in water, and (B) 80% acetonitrile, 0.08% formic acid. Peptides were eluted from the column with the following gradient 5% to 40% B (120 min), 40% to 80% (10 minutes). At 131 min, the gradient returned to 5% to re-equilibrate the column for 30 minutes before the next injection. Two blanks, one short 40 min-step gradient and one 60 min-linear gradient, were run between triplicates to prevent sample carryover. Peptides eluting from the column were analyzed by data dependent MS/MS, using top-10 acquisition method. Briefly, the instrument settings were as follows: resolution was set to 70,000 for MS scans and 17,500 for the data dependent MS/MS scans in order to increase speed. The MS AGC target was set to 3.106counts, while MS/MS AGC target was set to 5.104. The MS scan range was from 400 to 2000 m/z. MS and MS/MS scans were recorded in profile mode. Dynamic exclusion was set to 20 sec duration. Each sample was analyzed in biological triplicate by nanoLC/MS/MS. Each biological replicate was acquired in three technical replicates.PrideData Processing Following LC-MS/MS acquisition The MS files were processed with the MaxQuant software version 1.4.1.2 and searched with Andromeda search engine against the NCBI F. tularensis subsp. novicida database (release 28-04-2014; 1,719 entries). To search parent mass and fragment ions, we set an initial mass deviation of 4.5 ppm and 0.5 Da respectively, no match between runs allowed. The minimum peptide length was set to 7 amino acids and strict specificity for trypsin cleavage was required, allowing up to two missed cleavage sites. Carbamidomethylation (Cys) was set as fixed modification, whereas oxidation (Met) and N-term acetylation were set as variable modifications. The false discovery rates (FDRs) at the protein and peptide level were set to 1%. Scores were calculated in MaxQuant as described previously (Cox J., Mann M., 2008). The reverse and common contaminants hits were removed from MaxQuant output. Proteins were quantified according to the MaxQuant label-free algorithm using LFQ intensities [Luber, 2010 #1907;Cox, 2008 #1906]; protein quantification was obtained using at least 2 peptides (unique peptide + razor) per protein. Statistical and bioinformatic analysis, including heatmaps, profile plots and clustering, were performed with Perseus software (version 1.4.1.3) freely available at www.perseus-framework.org. For statistical comparison, we set two groups, each containing biological and technical triplicates, we then filtered the data to keep only proteins with at least 9 valid values out of 18. Next, the data were imputed to fill missing data points by creating a Gaussian distribution of random numbers with a standard deviation of 33% relative to the standard deviation of the measured values and 1.8 standard deviation downshift of the mean to simulate the distribution of low signal values. We performed a T-test, with a permutation based testing correction that was controlled by using a false discovery rate threshold of 0.05, 250 randomizations. Hierarchical clustering of proteins that survived the test was performed in Perseus on logarithmised LFQ intensities after z-score normalization of the data, using Euclidean distances using default parameters and setting 12 clusters for the row tree.ProteomicsIda Chiara GUERRERANecker proteomic platform, , PPN-3P5, Structure Federative de Recherche Necker, INSERM US24,PARTIALFrancisella Tularensis Subsp. Novicida (strain U112)chiara.guerrera@inserm.fr25616868 Ramond E, Gesbert G, Guerrera IC, Chhuon C, Dupuis M, Rigard M, Henry T, Barel M, Charbit A. Importance of host cell arginine uptake in Francisella phagosomal escape and ribosomal protein amounts. Mol Cell Proteomics. 2015 Jan 23. pii: mcp.M114.044552BiologicalNecker proteomics, INSERMFranceUpon entry into mammalian host cells, the pathogenic bacterium Francisella must import host cell arginine to multiply actively in the host cytoplasm. We identified and functionally characterized an arginine transporter (hereafter designated ArgP) whose inactivation considerably delayed bacterial phagosomal escape and intracellular multiplication. Intramacrophagic growth of the ΔargP mutant was fully restored upon supplementation of the growth medium with excess arginine, in both F. tularensis subsp. novicida and F. tularensis subsp. holarctica LVS, demonstrating the importance of arginine acquisition in these two subspecies. High-resolution mass spectrometry revealed that arginine limitation reduced the amount of most of the ribosomal proteins in the ΔargP mutant. In response to stresses such as nutritional limitation, repression of ribosomal protein synthesis has been observed in all kingdoms of life. Arginine availability may thus contribute to the sensing of the intracellular stage of the pathogen and to trigger phagosomal egress. All MS data have been deposited in the ProteomeXchange database with identifier PXD001584 (http://proteomecentral.proteomexchange.org/dataset/PXD001584).Importance of host cell arginine uptake in Francisella phagosomal escape and ribosomal protein amounts.Ramond Elodie E, Gesbert Gael G, Guerrera Ida Chiara IC, Chhuon Cerina C, Dupuis Marion M, Rigard Mélanie M, Henry Thomas T, Barel Monique M, Charbit Alain Asodium salt, liquid chromatography tandem mass spectroscopy, Water, IPP2A2, ammonium formate, 13C-labeled, MeCN, cadmium salt, Meth, NCMe, determination, CDM, DL-Arginine Acetate, postnatal development, instrument configuration, classic hairy cell leukaemia, H(2)O, zinc formate, Monohydrate, lead salt, BOUND WATER, growth and development, protein, Hydrogenchlorid, magnesium formate, oxidane, zinc salt, Solvent, WATER, HOH, 5730420M11Rik, 2-Propanone, Polypeptides, Techniques, ammonium tetraformate, hydrogen chloride, peptido, cobalt(II) formate dihydrate, Method, CH3-C#N, GRP1, Grp1, B1, SEC, Min, Arg, protein aggregate, Injectables, methyl ketone, DmelCG42273, DmelCG7212, SET, (S)-2-amino-5-guanidinopentanoic acid, Divorced, Eubacteria, LCMSMS, peptides, TAF-I, PTPSTEP, (S)-2-Amino-5-guanidinovaleric acid, sec, R, min, Arginine, HardwareType, mAPC, Monohydrate DL-Arginine Acetate, Divorces, copper, SCAN, free, NAG22, Bacteria Woese et al. 2024, AI047805, DmelCG4299, IGAAD, set, Wasserstoffchlorid, lithium salt, Bacteria <bacteria>, column, Methodological Studies, Propanon, DmelCG10574, Ice, Prokaryotae, l(2)k08110, D1, sample, dihydridooxygen, Strains, Procaryotae, dihydrogen oxide, ammonium (2:1) salt, nickel salt, GPH, HCAD, L-Arginin, LC-MS2, phapii, 6C6-AG, cobalt (+2) salt, dimethylketone, Arginine Hydrochloride, LC-MS/MS, aqua, Dmel_CG7826, arginine, StF-IT-1, Th, classic hairy cell leukemia, Procedure, Arginin, Thermo Scientific, Aceton, shortened, H2O, sodium (4:1:1) salt, Pyroacetic ether, Dm1, magnesium salt, H-CAD, 3.1.3.48, Chlorwasserstoff, prokaryotes, nickel (+2) salt, Dmel_CG7835, Hydrogen Oxide, Crystal, L-Arginine, Mnb, MNB, DXS1357E, ammonium salt, HLA-DR-associated protein II, growth pattern, instrument, DI-2, Step, CG11628, non-developmental growth, I-2Dm, HCl, formate, cobaltous formate, hydrogen hydroxide, cromium (+3), Striatum-enriched protein-tyrosine phosphatase, Methodological, CG4299, AW124434, beta-Ketopropane, beta Trypsin, Methodological Study, acqua, I-2PP1, L-CAD, Speed, biological replicate, TAF-IBETA, STEP, label, liquid chromatography-tandem mass spectroscopy, copper salt, ammonium (4:1) salt, TAF-Ibeta, Polypeptide, Wasser, CG7212, methyl cyanide, chlorane, Imp13, lead (+2) salt, short, Strains and Sprains, DDCH, i2pp2a, Prokaryota, nickel formate dihydrate, GRP1/cytohesin 1, Procedures, not genetically inherited., lead formate, chlorure d'hydrogene, aluminum salt, cesium salt, number, 2-Amino-5-guanidinovaleric acid, L Isomer, Gene, imp13, eubacteria, 3R23, protein-containing complex, thomson, Chalk, presence, CG7826, PHAPII, LC-MS-MS, agua, hardware, L-(+)-arginine, formic acid, mass-to-charge ratio, CG11633, cytohesin/GRP1, method, propanone, potassium salt, (2S)-2-amino-5-(carbamimidamido)pentanoic acid, biological_replicate, Injection, Harg, 14C-labeled, method used in an experiment, LC-MSMS, dimethylcetone, Tina, Gene Products, CG7835, Studies, CG42273, Bacteriobiota, Sprains, Separated, cloruro de hidrogeno, Technique, CG 7212, Pro-Mega, T6K12_14, Glass, Hydrogen chloride, thallium (+1) salt, Selb, ipp2a2, beta-Trypsin, rubidium salt, stubby, l(2)SH2 0323, 2pp2a, eau, calcium formate, methanoic acid, Monera, CG10574, Bacteria (ex Cavalier-Smith 1987), Injectable, Study, Neural-specific protein-tyrosine phosphatase, 2PP2A, Suspension, taf-ibeta, dSET, dSet, L-Arg, peptidos, cromium (+3) salt, liquid, fungi, Azeton, Dimethyl ketone, Chromatographies, 2-amino-5-(carbamimidamido)pentanoic acid, sodium formate, PTHB1, bacteria, DYRK1, Dimethylketon, 1-(14)C-labeled, nickel formate, 3H-labeled, protein complex, sd-5, Proteins, Sprain, igaad, strontium formate, stepk, chloridohydrogen, potassium formate, 2-amino-5-guanidinopentanoic acid, copper (+2) salt, backward, L-Isomer, acetonitrile, strontium salt, DL Arginine Acetate, Cell, Peptide, group, development, [HCl], count in organism, LC/MS/MS, secret agent, native protein, L-Isomer Arginine, I-2PP2A, C18, cupric formate, water, Protein, chemical analysis, Dm I-2, Strain, I2PP2A, Hydrochloride, L Arginine, Dyrk1, Vacuums, aluminum formate, l(2)SH0323, ACETONITRILE, cyanomethane, T6K12.14, CYH1, AU020952, Separation, lithium formate, PRSS, ensemble, BAP31, Separations, (2S)-2-amino-5-guanidinopentanoic acid, CC1, postnatal growth, Tripcellim, ethanenitrile, chromic formate, sample population, DmelCG11628, ME-IV, Protein Gene Products, plan specification, Gene Proteins, Trypure, dSET/TAF-Ibeta, 2610030F17Rik, concentration, prokaryote, calcium salt, liquid chromatography tandem mass spectrometry, Peptid, assay, [OH2], LCAD, AA407739, growth, hydrochloric acid, reversedL-Arginine, Peptidomics., (S)-2-amino-5-guanidinopentanoic acid, DL-Arginine Acetate, (2S)-2-amino-5-guanidinopentanoic acid, (S)-2-Amino-5-guanidinovaleric acid, Arginine Hydrochloride, 2-Amino-5-guanidinovaleric acid, R, L Isomer, arginine, Monohydrate, Arginine, Monohydrate DL-Arginine Acetate, 2-amino-5-guanidinopentanoic acid, Arginin, L-Isomer, DL Arginine Acetate, L-(+)-arginine, Francisellaceae, (2S)-2-amino-5-(carbamimidamido)pentanoic acid, L-Isomer Arginine, Harg, Hydrochloride, L-Arg, L Arginine, Arg, 2-amino-5-(carbamimidamido)pentanoic acid, L-Argininliquid chromatography tandem mass spectroscopy, IPP2A2, MRG-2, cox4, E 920, ion, Hgfr, determination, supply, Aminosaeure, cleavage, Amino acid, 5730555F13Rik, protein, 5730420M11Rik, peptide, Polypeptides, peptido, HGFR, Memory-related gene 2 protein, Software Engineering, 2-amino-3-mercaptopropanoic acid, Cystein, DmelCG10664, protein aggregate, Gpi-1, Computer Program, WMS, SET, C, me75, F, LCMSMS, amino acids, peptides, reference sample, TAF-I, MUB3_18, cisteina, COXIV, Nlk, M, Open, Computer Programs and Programming, GDH 1, GDH 2, MUB3.18, COXA_DROME, free, D17Mit170, T1, DmelCG4299, IGAAD, set, Cox4, DmelCG10574, ppm, 1.3.3.3, RG7MT1, GPHYSD2, Racemethionine, COX-VA, SGS, NK/GPI, LC-MS2, phapii, Gpi, proto-oncogene c-Met, PRO, Aminokarbonsaeure, acetylation, LC-MS/MS, 2-Amino-4-(methylthio)butyric acid, StF-IT-1, Search, 9130215G10Rik, Tl3, Tl2, CYS, Cys, Source Softwares, alpha-amino carboxylic acids, Software Tools, Programs, ACMICD, Program, Computer Applications, mOC-X, Ionen, VA, AL024441, Algorithm, Computer Applications Software, Computer Applications Softwares, Methionin, NK|GPI, Softwares, Hmet, COX IV, (2R)-2-amino-3-sulfanylpropanoic acid, and GLY protein 2, HGF receptor, HGF/SF receptor, Amino Acid, Acid, oligonucleotide random primer, and GLY protein 1, Org, ORG, Software Applications, HLA-DR-associated protein II, Amino acids, DI-2, Va, Coprogen oxidase, Source Software, I-2Dm, COX IV-1, common, Gpi1-r, Gpi1-s, MASS, CG4299, hCMT1c, c-Met, Gpi1-t, beta Trypsin, L-Cystein, I-2PP1, data analysis, Applications, Tree., TAF-IBETA, liquid chromatography-tandem mass spectroscopy, label, TAF-Ibeta, Acids, Polypeptide, DmelCG14724, i2pp2a, Bglap-rs1, Cox4a, Gpi1s, Computer Software Applications, 2-amino-4-(methylsulfanyl)butanoic acid, False, AI461847, Aminocarbonsaeure, L-Zystein, FBN, Gene, Computer, alpha-amino acid, protein-containing complex, PHAPII, LC-MS-MS, CG10664, HGF, ECTOL1, IV, LC-MSMS, Gene Products, F15E12.6, Pgi, Low, RCCP2, Application, Open Source Softwares, Search Engines, F15E12_6, Coproporphyrinogenase, scatter factor receptor, Gpi-1r, distribution, Software Application, iones, Gpi-1s, E-920, ipp2a2, Phi, beta-Trypsin, Gpi-1t, Open Source Software, Acetylations, 2pp2a, Hcys, ions, CG10574, AUTS9, Computer Software Application, OCTD, RANDOM, data processing, 2PP2A, 2-amino-4-(methylthio)butanoic acid, Tools, 10^[-6], CYSTEINE, taf-ibeta, COX, dSET, dSet, FREE CYSTEINE, peptidos, AI838057, (2R)-2-amino-3-mercaptopropanoic acid, metionina, 1.4.1.3, Controlled, Applications Software, Open Source, Controlling, Computer Software, cou, DL-Methionine, protein complex, Proteins, igaad, alpha-amino acids, MF, Amf, Peptide, Engine, Tool, group, polypeptide, LC/MS/MS, Software Tool, Lr, native protein, I-2PP2A, Dm I-2, Protein, chemical analysis, I2PP2A, MFS1, IV-1, supply and distribution, Software, Zystein, Data Base, WMS2, ATCMPG1, MET, Met, NK, ATCMPG2, Random selection by shearing, PRSS, ensemble, 2-amino-3-sulfanylpropanoic acid, tyrosine-protein kinase Met, Engineering, tend, Tripcellim, L-2-Amino-3-mercaptopropionic acid, (R)-2-amino-3-mercaptopropanoic acid, Amino, 2-Amino-3-mercaptopropionic acid, Protein Gene Products, Computer Programs, Trypure, Gene Proteins, dSET/TAF-Ibeta, 2610030F17Rik, CG14724, Applications Softwares, Ion, SSKS, alpha-amino-gamma-methylmercaptobutyric acid, Bra, liquid chromatography tandem mass spectrometry, E920, Peptid, assay, Par4, variable, AA407739, 5730455C01Rik, SF receptorP-LAP, MGC130048, host organism, Dendritic Cells, Zoonotic Infection, nucleocytoplasm, Zoonotic Infectious, Laboratory, DL-Arginine Acetate, Interdigitating, Zoonotic Infectious Disease, Aminosaeure, Mus domesticus, Amino acid, A4, Monohydrate, ribosomal RNA, Ribosomal Protein, CASP-14, Stage, Life History, House Mouse, Social Controls, Membrane Tissues, Monocyte-Derived Macrophage, Life Cycle, Interdigitating Dendritic Cell, Arg, Dendritic Cell, Infectious Diseases, phagosome, Interstitial Dendritic Cell, Formal Social Controls, gamma sarcoglycan, Phagolysosomes, Ribosomal, (S)-2-amino-5-guanidinopentanoic acid, Eubacteria, amino acids, Phagosome, entire life cycle, (S)-2-Amino-5-guanidinovaleric acid, membrane region, Tissue, R, Plasmacytoid, hypoplasia, Arginine, Swiss Mice, Monohydrate DL-Arginine Acetate, Francisella tularense, Bacteria Woese et al. 2024, Social, Bacteria <bacteria>, Oxytocinase, Prokaryotae, gamma-sarcoglycan, 4732490P18Rik, stage, Procaryotae, Pahvant Valley plague, Francisella tularensis, L-Arginin, Interstitial Dendritic Cells, Membrane Tissue, Infectious Disease, Aminokarbonsaeure, Arginine Hydrochloride, CASP14, mouse, SG-gamma, arginine, integral to membrane, Arginin, Interstitial, alpha-amino carboxylic acids, Francisellaceae, Pasteurella tularensis, Plasmacytoid Dendritic Cells, Mini-ICE, Diseases, integral component of membrane., sarcoglycan, prokaryotes, internal to cell, Monocyte-Derived Macrophages, Insulin-regulated membrane aminopeptidase, L-Arginine, Amino Acid, Acid, Mus musculus, Amino acids, Caspase-14 subunit p10, ribosomal protein, mice, life, Life Cycle Stage, Veiled, Swiss Mouse, lemming fever, Control, History Stage, Caspase-14 subunit p19, rabbit fever, Controls, MICE, gamma (35kDa dystrophin-associated glycoprotein), domesticus, Life Cycles, DMDA, 35kD dystrophin-associated glycoprotein, Cells, OTase, Interdigitating Dendritic Cells, Acids, Mouse, Regulation, gp160, Prokaryota, 2010309L07Rik, Regulations, 3.4.22.-, vp165, lifespan, SGCG_HUMAN, Monocyte Derived Macrophages, PLAP, developmental stage, Aminocarbonsaeure, 2-Amino-5-guanidinovaleric acid, L Isomer, Gene, mini-ICE, Yatobyo (Japan), eubacteria, alpha-amino acid, Zoonotic Infectious Diseases, Francisella tularensis infection, Deerfly fever, Zoonotic, TYPE, Zoonotic Spillovers, L-(+)-arginine, DAGA4, Zoonotic Infections, (2S)-2-amino-5-(carbamimidamido)pentanoic acid, reduced, House, Bacterium tularense, Harg, Cycle, GP160, 35DAG, Gene Products, Tularemias, Mus musculus domesticus, Vesicle protein of 165 kDa, Bacteriobiota, tiny, Life, MAM, gamma-SG, IRAP, Mice, SCG3, protoplasm, Cycles, Spillovers, protoplast, Infections, Plasmacytoid Dendritic, entire lifespan, Brucella tularensis, Swiss, Tissues, Interdigitating Dendritic, Dendritic, Phagolysosome, Bone Marrow-Derived Macrophage, Life History Stages, Placental leucine aminopeptidase, Monera, Insulin-responsive aminopeptidase, Bone Marrow-Derived, Bacteria (ex Cavalier-Smith 1987), Interstitial Dendritic, Stages, Life History Stage, region of membrane, L-Arg, fungi, Cystinyl aminopeptidase, Vp165, 2-amino-5-(carbamimidamido)pentanoic acid, small, membrane, Disease, Veiled Cells, bacteria, 35 kDa dystrophin-associated glycoprotein, Formal Social Control, Francisella tularensis Infection, Proteins, alpha-amino acids, Monocyte Derived, 2-amino-5-guanidinopentanoic acid, L-Isomer, DL Arginine Acetate, Cell, SGCG, LGMD2C, Zoonotic Spillover, Monocyte-Derived, uptake, L-Isomer Arginine, Mus, Social Control, Protein, Infection, Francisella tularensis Infections, Hydrochloride, whole membrane, CAP, L Arginine, Ohara disease, Macrophage, transmembrane, Plasmacytoid Dendritic Cell, History Stages, DMDA1, Veiled Cell, underdeveloped, Macrophages, (2S)-2-amino-5-guanidinopentanoic acid, Interdigitating Cells, Zoonotic Diseases, Bone Marrow Derived Macrophages, House Mice, Amino, Membrane, Laboratory Mice, Protein Gene Products, Gene Proteins, prokaryote, SCARMD2, Bone Marrow-Derived Macrophages, 3.4.11.3, Zoonotic Disease, Interdigitating Cell, regulation, Laboratory Mousehost organism, lifespan, nucleocytoplasm, AI461847, DL-Arginine Acetate, postnatal development, developmental stage, 2-Amino-5-guanidinovaleric acid, L Isomer, Monohydrate, ribosomal RNA, Ribosomal Protein, growth and development, Spectrum Analyses, Protoplasm, L-(+)-arginine, (2S)-2-amino-5-(carbamimidamido)pentanoic acid, reduced, Harg, Mass, Pgi, Analysis, tiny, Arg, Gpi-1, Mass Spectroscopy, protoplasm, Mass Spectrum Analysis, Ribosomal, (S)-2-amino-5-guanidinopentanoic acid, protoplast, Analyses, availability, entire lifespan, Gpi-1r, Nlk, entire life cycle, Gpi-1s, (S)-2-Amino-5-guanidinovaleric acid, Phi, R, hypoplasia, Gpi-1t, Arginine, Monohydrate DL-Arginine Acetate, L-Arg, stage, 2-amino-5-(carbamimidamido)pentanoic acid, L-Arginin, NK/GPI, small, Data Set., Gpi, Cytoplasms, Proteins, Arginine Hydrochloride, arginine, 2-amino-5-guanidinopentanoic acid, MF, Arginin, L-Isomer, DL Arginine Acetate, Amf, Spectrum Analysis, Cell, Spectroscopy, Francisellaceae, development, MS, mOC-X, L-Isomer Arginine, Protein, Hydrochloride, L Arginine, NK|GPI, internal to cell, Mass Spectrum Analyses, Data Base, NK, L-Arginine, Mass Spectrum, Org, ORG, growth pattern, underdeveloped, ribosomal protein, non-developmental growth, life, (2S)-2-amino-5-guanidinopentanoic acid, postnatal growth, Spectrometry, Gpi1-r, Gpi1-s, Gpi1-t, Protoplasms, growth, Bglap-rs1, Gpi1sL-Arginine, Ribosomal, (S)-2-amino-5-guanidinopentanoic acid, DL-Arginine Acetate, (2S)-2-amino-5-guanidinopentanoic acid, (S)-2-Amino-5-guanidinovaleric acid, Proteins, Arginine Hydrochloride, 2-Amino-5-guanidinovaleric acid, R, L Isomer, arginine, Monohydrate, Arginine, ribosomal RNA, Ribosomal Protein, Monohydrate DL-Arginine Acetate, 2-amino-5-guanidinopentanoic acid, Arginin, L-Isomer, DL Arginine Acetate, ribosomal protein., L-(+)-arginine, Francisellaceae, uptake, (2S)-2-amino-5-(carbamimidamido)pentanoic acid, L-Isomer Arginine, Harg, Protein, Hydrochloride, L-Arg, L Arginine, Arg, 2-amino-5-(carbamimidamido)pentanoic acid, L-Arginin332falseImportance of arginine trasporter ArgP in Francisella by comparative proteomics.Metabolic adaptation has recently been recognized as a key component of the intracellular life cycle of pathogenic microorganisms. Upon entry into mammalian host cells, Francisella tularensis, a pathogenic bacterium responsible for the zoonotic disease tularemia, rapidly escapes from the phagosome to multiply actively in the cytosolic compartment. Francisella is auxotroph for several amino acids, including arginine, and thus relies exclusively on their import from the host to survive in infected cells. We identified an arginine transporter (hereafter designated ArgP) whose inactivation considerably delayed bacterial phagosomal escape and intracellular multiplication. Remarkably, this defect could be totally suppressed in macrophages and dendritic cells from IRAP-KO mice. Wild-type cytosolic multiplication of the mutant bacteria was restored in these cells, indicating that the ArgP transporter is mainly required for arginine uptake in the phagosomal compartment. Proteomic analyses showed that arginine deprivation in the ∆argP mutant rapidly affected the expression of a numerous proteins, and in particular reduced expression of most of the ribosomal proteins. We propose that regulation of ribosomal protein biogenesis may contribute to the sensing of the intracellular stage of the pathogen and to phagosomal membrane disruption.2015-01-282015-01-13PXD001584NEWT:68239NEWT:108018NEWT:377960NEWT:1131NEWT:2042546NEWT:148942NEWT:295546NEWT:258594NEWT:1126NEWT:1129NEWT:309807NEWT:89184NEWT:309800NEWT:1323429NEWT:876138NEWT:44271NEWT:193516NEWT:2315688NEWT:592321NEWT:1073882NEWT:6819NEWT:428386NEWT:1117NEWT:498257NEWT:10036NEWT:32264NEWT:10034NEWT:661410NEWT:224326NEWT:376619NEWT:3765NEWT:3767NEWT:433727NEWT:135622NEWT:672127NEWT:656061NEWT:37965NEWT:656064NEWT:3760NEWT:36630NEWT:381198NEWT:418106NEWT:10029NEWT:913645NEWT:2492962NEWT:641809NEWT:44250NEWT:317447NEWT:5530NEWT:868565NEWT:135674NEWT:1562698NEWT:12637NEWT:2164133NEWT:2767846NEWT:314344NEWT:108061NEWT:224308NEWT:49928NEWT:160621NEWT:110368NEWT:1916073NEWT:1310161NEWT:1230383NEWT:77133NEWT:1310165NEWT:114727NEWT:5516NEWT:498211NEWT:347256NEWT:5518NEWT:398007NEWT:1527468NEWT:498217NEWT:498216NEWT:11320NEWT:1310173NEWT:37919NEWT:161934NEWT:156587NEWT:1148NEWT:5508NEWT:419481NEWT:5507NEWT:1140NCBITaxon:2157NEWT:2066563NEWT:1143NEWT:1462472NEWT:1336795NEWT:644042NEWT:709964NEWT:3712NEWT:3711NEWT:3714NEWT:3715NEWT:2065263NEWT:1549675NEWT:177437NEWT:9014NEWT:10891NEWT:226126NEWT:1616117NEWT:34865NEWT:28174NEWT:550537NEWT:284812NEWT:3701NEWT:3702NEWT:1245466NEWT:3704NEWT:1246791NEWT:9005NEWT:34871NEWT:9006NEWT:33548NEWT:1883426NEWT:657321NEWT:10886NEWT:3708NEWT:536231NEWT:219813NEWT:215451NEWT:460519NEWT:272844NEWT:1348799NEWT:55784NEWT:485NEWT:2709072NEWT:2499525NEWT:142786NEWT:2853422NEWT:487NEWT:1679718NEWT:480NEWT:67767NEWT:46835NEWT:216778NEWT:376686NEWT:237610EFO:0001352NEWT:219809NEWT:219808NEWT:119562NEWT:1883446NEWT:1233435NEWT:118223NEWT:141469NEWT:987059NEWT:491NEWT:460523NEWT:85694NEWT:85692NEWT:3750NEWT:291146NEWT:29466NEWT:3745NEWT:1715989NCBITaxon:4751NEWT:3747NEWT:278161NEWT:415978NEWT:410289NEWT:191701NEWT:9054NEWT:472NEWT:29459NEWT:1380096NEWT:28128NEWT:470NEWT:41122NEWT:1328300NEWT:748693NCBITaxon:50557NEWT:3735NEWT:29491NEWT:911022NEWT:446NEWT:153481NEWT:445NEWT:2003758NEWT:63366NEWT:63367NEWT:29488NEWT:215402NEWT:3726NEWT:30455NEWT:28141NEWT:9030NEWT:9031NEWT:3728NEWT:108931NEWT:219799NEWT:219798NEWT:548NEWT:150475NEWT:256737NEWT:51350NEWT:8694NEWT:8692NEWT:542NEWT:394503NEWT:13076NEWT:219785NEWT:451515NEWT:8673NEWT:550NEWT:10804NEWT:554NEWT:451516NEWT:552NEWT:1325291NEWT:42410NEWT:1225786NEWT:771870NEWT:8665NEWT:51338NEWT:9995NEWT:28112NEWT:49012NEWT:28110NEWT:10832NEWT:507601NEWT:520NEWT:64657NEWT:186441NEWT:2585209NEWT:643680NEWT:214092NEWT:40697NEWT:13095NEWT:162425NEWT:242124NEWT:9986NEWT:8654NEWT:8658NEWT:1268063NEWT:8657NEWT:9983NEWT:8655NEWT:595495NEWT:80833NEWT:479096NEWT:28108NEWT:28104NEWT:1114957NEWT:568703NEWT:568708NEWT:52641NEWT:353152NEWT:353153NEWT:227377NEWT:1774284NEWT:27689NEWT:207769NEWT:40674NEWT:51329NCBITaxon:1763NEWT:443906NEWT:13037NEWT:519NEWT:7394NEWT:6063NEWT:404330NEWT:1328388NEWT:300641NEWT:39655NEWT:38323NEWT:206411NEWT:80869NEWT:214053NEWT:80863NEWT:665890NEWT:52639NEWT:52638NEWT:8697NEWT:861109NEWT:8695NEWT:1196866NEWT:510516NEWT:1045775NEWT:1123869NCBITaxon:1335626NEWT:7370NEWT:83906NEWT:99488NEWT:1134506NEWT:255470NEWT:27697NEWT:86185NEWT:9935NEWT:287889NEWT:1028307NEWT:52603NEWT:92867NEWT:92866NEWT:1344414NEWT:202950NEWT:741158NEWT:1293091NEWT:202956NEWT:315750NEWT:632957NEWT:9925NCBITaxon:9606NEWT:1436183NEWT:4232NEWT:9922NEWT:416348NEWT:105812NEWT:11298NEWT:4236NEWT:398031NEWT:196627NEWT:523840NEWT:1076878NEWT:200308NEWT:200302NEWT:9913NEWT:9915NEWT:105841NEWT:1170227NEWT:2666255NEWT:999810NEWT:28995NEWT:1392998NEWT:1150621NEWT:1085791NEWT:745531NEWT:226900NEWT:231490NEWT:590651NEWT:449239NEWT:244704NEWT:9909NEWT:9902NEWT:430615NEWT:563041NEWT:36599NEWT:41949NEWT:9900NEWT:326423NEWT:326424NEWT:13013NEWT:749906NEWT:418985NEWT:749907NEWT:431947NEWT:142809NEWT:1095183NEWT:302409NEWT:9979NEWT:130821NEWT:2320211NEWT:27606NEWT:290610NEWT:1519788NEWT:59202NEWT:9975NEWT:8643NEWT:1801631NEWT:1159899NEWT:502780NEWT:860688NEWT:35231NEWT:1774208NEWT:8644NEWT:1125630NEWT:28930NEWT:202914NEWT:222523NEWT:27602NEWT:502779NEWT:297246NEWT:483139NEWT:101491NEWT:8639NEWT:8637NEWT:1114970NEWT:285272NEWT:192222NEWT:9963NEWT:197911NEWT:197912NEWT:239386NEWT:2785785NEWT:82633NEWT:1298722NEWT:8626NEWT:105884NCBITaxon:9615NEWT:8621NEWT:8620NEWT:8624NEWT:8623NEWT:5597NEWT:218496NEWT:767769NEWT:8618NEWT:70448NEWT:9948NEWT:1088878NEWT:9945NEWT:9940NEWT:8612NEWT:315749NEWT:1247190NEWT:2126719NEWT:555479NEWT:1079921NEWT:1145230NEWT:1538089NEWT:6945NEWT:2627626NEWT:6943NEWT:870726NEWT:91374NEWT:91373NEWT:27804NEWT:474186NEWT:10136NEWT:638301NEWT:94400NEWT:6941NEWT:307972NEWT:755969NEWT:693153NEWT:34NEWT:200492NEWT:1299114NEWT:44386NEWT:1590841NEWT:1276473NEWT:2282334NEWT:1246NEWT:1247NEWT:70667NEWT:869728NEWT:6938NEWT:1885322NEWT:36774NEWT:194963NEWT:77298NEWT:15819NEWT:109461NEWT:1085956NEWT:1236NEWT:1136157NEWT:6928NEWT:1239NEWT:36745NEWT:882735NEWT:400727NEWT:90062NEWT:83772NEWT:68335NEWT:332952NEWT:3885NEWT:3888NEWT:2653900NEWT:32391NEWT:6915NEWT:101510NEWT:3880NEWT:216816NEWT:1220NEWT:10140NEWT:10141NEWT:95767NEWT:466585NEWT:29NEWT:6918NEWT: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