{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Tabular":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD070949/report.tsv"],"Txt":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD070949/checksum.txt"],"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD070949/EG_Prt-17-2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD070949/EG_Prt-17-3.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD070949/EG_Prt-17-4.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD070949/EG_Prt-17-5.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD070949/EG_Prt-17-1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD070949/EG_Prt-17-6.raw"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["timo.glatter@mpi-marburg.mpg.de"],"submitter":["Timo Glatter"],"technology_type":["Mass Spectrometry","Bottom-up proteomics","Data-independent acquisition"],"software":[""],"submitter_keywords":["T5-like phage","E.coli"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD070949"],"sample_protocol":["For proteome analyses, samples were takenby harvesting 2 ml cells at 16,000 g and 4°C for 2 min and subsequent freezing of cell pellets in liquid nitrogen. In order to prepare samples for proteome measurements, cells were thawed on ice and lysed by resuspending in 200 µL of lysis buffer (2% sodium lauroyl sarcosinate (SLS) in 100 mM ammonium bicarbonate (ABC) buffer), heating at 90°C for 15 min and 3 cycles of sonication (30s each) using a Hielscher Up200St ultrasonic processor (75% amplitude, 0.5 s / 0.5 s pulse) . Subsequently, samples were centrifuged at 14,000 rpm for 5 min and supernatants were transferred into a new low protein binding tube and protein concentrations were determined using a PierceTM BCA Protein assay kit (ThermoFisher Scientific, 23250). Afterwards, 2 mM final tris(2-carboxyethyl)phosphine (TCEP, Tokyo Chemical Industry, ref. T1656) were added and samples were mixed and incubated at 90°C for 15 min to promote reduction of disulfide bridges. After cooling down, 4 mM iodacetamide were added and samples were incubated in the dark for 30 min at room temperature. Afterwards, 30 µg of total lysate proteins were transferred to a new low protein binding tube and sample volume was adjusted with lysis buffer to a total volume of 50 µL. Then, 3 volumes of 100 mM ABC buffer were added tot he sample in order to dilute the final SLS concentration down to 0.5%. Then, SP3 magnetic beads slurry was prepared as follow: in a new tube, 20 µL of beads (Sera-MagTM carboxylate-modified [E3] and [E7] magnetic beads, Cytiva) were mixed at a 1:1 ratio and washed twice with 300 µL of HPLC grade water. Then, beads were resuspended in 100 µL of water to produce readily usable slurry. Afterwards, 8 µL of bead slurry was added to the protein sample along with 3 sample volumes (600 µl) of 100% acetonitrile (ACN). Proteins were allowed to bind to the beads for 1 h at RT with 800 rpm shaking. Supernatant was discarded by using a magentic rack and beads were washed twice with 300 µl of 70% ethanol, once with 300 µL of 100% ACN before air drying for 5 min. Finally, protein digest was performed by adding 200 µL of 100 mM ABC buffer and 0.6 µg of sequencing-grade trypsin (Promega, V5111) overnight at 30°C with 1,000 rpm shaking. Supernatants were transferred to a new clean low protein binding tube and beads were resuspended in 100 µL of ddH2O. After magnetic bead separation, supernatants were pooled with the previous ones and residual SLS was precipitated by addition of 4% final concentration of trifluoroacetic acid (TFA, Serva 45641.01). Samples were centrifuged at 4°C at 14,000 rpm for 20 min and supernatants were purified with C18 columns (Chromabond, 730522.250). The solvent was evaporated with SpeedVac vacuum concentrations at 45°C and stored samples at -20°C before measurement.  Liquid Chromatography-Mass Spectrometry (LC-MS) for peptide analysis was carried out using a Vanquish Neo system coupled to an Orbitrap Exploris 480 mass spectrometer. Peptides were dissolved 0.1% trifluoroacetic acid and loaded onto a self-packed C18 (26 cm of 1.9 μm Reprosil-AQ, Dr. Maisch) column. The peptides were separated by an acetonitrile gradient running from 2 – 25 % solvent B (99.85% acetonitrile, 0.15% formic acid) over 45 min, followed by an additional increase of  solvent B to 35% and 40 %, respectively, for 15 min at a flow rate of 300 nl/min. Solvent A composition was 0.15 % formic acid.  Eluting peptides were analyzed in data independent acquisition (DIA) mode on the Exploris-MS. The funnel RF level was set to 40. Full MS resolution was set to 120.000 (m/z 200). Automatic gain control (AGC) target value for fragment spectra was set at 3000%. 45 windows of 14 Da were used with an overlap of 1 Da between m/z 320-950. Resolution was set to 15,000 and fill time to 22 ms. Stepped HCD collision energy of 25, 27.5, 30 % was used. MS1 data was acquired in profile, MS2 DIA data in centroid mode."],"repository":["Pride"],"modification":[""],"quantification_method":["Not available"],"data_protocol":["Analysis of DIA data was performed using the DIA-NN version 1.9 (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6949130/), respectively, using a uniprot protein database from E.coli with Phage proteins included to generate a data set specific spectral library for the DIA analysis. The neural network based DIA-NN suite performed noise interference correction (mass correction, RT prediction and precursor/fragment co-elution correlation) and peptide precursor signal extraction of the DIA-NN raw data. The following parameters were used: Full tryptic digest was allowed with two missed cleavage sites, and oxidized methionines (variable) and carbamidomethylated cysteins (fixed). Match between runs and remove likely interferences were enabled. The precursor FDR was set to 1%. The neural network classifier was set to the single-pass mode. Quantification strategy was set to any LC (“high accuracy” for DIA-NN 1.8) and Quant UMS (“high precision” for DIA-NN 1.9). Cross-run normalization was set to RT-dependent. Library generation was set to smart profiling. DIA-NN outputs were further evaluated using the SafeQuant (https://pubmed.ncbi.nlm.nih.gov/23017020/ & https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/pmic.201300135) script modified to process DIA-NN outputs."],"omics_type":["Proteomics"],"labhead":["Timo Glatter"],"instrument_platform":[""],"labhead_affiliation":["Max Planck Institute for Terrestrial Microbiology Karl-von-Frisch Str. 10  35043 Marburg Germany"],"submission_type":["PARTIAL"],"species":["Escherichia Coli","Escherichia Phage Hildybeyeler"],"publication":["Not available"],"submitter_mail":["timo.glatter@mpi-marburg.mpg.de"],"submitter_affiliation":["Max Planck Institute for Terrestrial Microbiology\nMarburg"],"submitter_country":["Germany"],"additional_accession":[]},"is_claimable":false,"name":"DNA-intercalating antiphage molecules trigger abortive infection through ‘mutual destruction’ and synergize with bacterial immunity","description":"Bacteria deploy diverse antiphage defense systems, including small bioactive molecules providing protection at the multicellular level. DNA-intercalating anthracyclines such as daunorubicin exhibit broad antiphage activity, but the underlying mechanism had remained elusive. Here, we systematically screened the Escherichia coli BASEL phage collection to elucidate the mode of action of daunorubicin. We identified taxonomically distinct clusters of susceptible viral groups and demonstrate that daunorubicin blocks infection of T5-like phages (Markadamsvirinae) after first-step transfer (FST) as revealed by long-read sequencing. Continued expression of pre-early genes leads to abortive infection via ‘mutual destruction’, where both phage and host succumb. Analogous phenotypes of abortive infection were observed for taxonomically diverse phages with different DNA-intercalating antiphage molecules. Notably, we show that daunorubicin synergizes with downstream nucleic acid-targeting defences underscoring context dependency in the observed defense phenotype. Our findings reveal how chemical defense contributes to the multilayered antiviral immunity and highlight the intricate interplay between mechanistic inhibition and infection outcome.","dates":{"publication":"2026-05-13","submission":"2025-11-19"},"accession":"PXD070949","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:1131","NEWT:2042546","NEWT:259447","NEWT:295546","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:1073882","NEWT:6819","NEWT:475174","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:1582","NEWT:1584","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:7719","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NCBITaxon:37162","NEWT:2065263","NEWT:177437","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2499525","NEWT:2853422","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:108458","NEWT:5936","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:5911","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:11723","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:906914","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:548","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:2126348","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:13076","NEWT:207340","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:550","NEWT:33941","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:9995","NEWT:575412","NEWT:28112","NEWT:49012","NEWT:6493","NEWT:10832","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:5147","NEWT:28108","NEWT:28104","NEWT:407821","NEWT:568703","NCBITaxon:2","NEWT:568708","NEWT:1613","NEWT:986","NEWT:52641","NEWT:28532","NEWT:353152","NEWT:298176","NEWT:353153","NEWT:227377","NEWT:1774284","NEWT:40674","NEWT:1194669","NEWT:74426","NEWT:443906","NEWT:13037","NEWT:519","NEWT:2510939","NEWT:7394","NEWT:6063","NEWT:1328388","NEWT:105487","NEWT:1548728","NEWT:667127","NEWT:9557","NEWT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