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Hay"],"technology_type":["Data-dependent acquisition","Mass Spectrometry","Bottom-up proteomics","Data-independent acquisition"],"software":[""],"submitter_keywords":[""],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD060209"],"sample_protocol":["Trifluoroethanol cell pellet lysis Cells were washed with PBS, scraped for collection, and pelleted in a microcentrifuge at maximum speed for 15 min at 4°C. Any remaining PBS was removed by micropipette and the pellet was flash frozen in liquid nitrogen. Cells were resuspended by vortexing (VWR) with 150 μL of 50% trifluoroethanol (TFE) followed by sonication (Fisher Scientific) for 5 min in an ice bath. Samples were reduced and alkylated for 10 min at 95°C in a solution of 100mM Tris pH 8.5, 10 mM tris(2-carboxyethyl)phosphine (TCEP, Bioshop), and 40 mM chloroacetamide (CAA, Acros Organics). Samples were diluted to at most 10% TFE with 50 mM ammonium bicarbonate. Lys-C/trypsin (Promega) was added at a 1:50 ratio of protease:protein and incubated overnight at 37°C, followed by additional lys-C/trypsin at a 1:200 ratio incubated for 6 h at 37°C. Following TFE cell pellet lysis and digestion, peptide samples were desalted and purified on C-18 STAGE tips as described.(Rappsilber, Ishihama and Mann, 2003)  LC-MS/MS (RPL28 knockdown) Purified peptides were analyzed using a quadrupole time of flight mass spectrometer (Impact II; Bruker Daltonics) on-line, coupled to an Easy nano LC 1200 HPLC (Thermo Fisher) using nanoBooster with methanol and a Captive spray nanospray ionization source (Bruker Daltonics). Buffer A was composed of 0.1% formic acid and 2% acetonitrile in H2O and buffer B was composed of 0.1% formic acid and 80% acetonitrile in H2O. Samples were resuspended in buffer A and 100 ng were loaded onto the instrument running a standard 90-minute gradient. The Impact II was set to data-dependent acquisition.  LC-MS/MS (BOP1 knockdown) Purified peptides were analyzed using a quadrupole time of flight mass spectrometer (timsTOF Pro; Bruker Daltonics) operated in DIA-PASEF mode, coupled to an NanoElute UHPLC system (Bruker Daltonics) with Aurora Series Gen2 (CSI) analytical column (25 cm x 75 μM 1.6 μM FSC C18, with Gen2 nanoZero and CSI fitting; Ion Opticks, Parkville, Victoria, Australia). Buffer A was composed of 0.1% formic acid and 0.5% acetonitrile in H2O and buffer B was composed of 0.1% formic acid in acetonitrile. Samples were resuspended in buffer A and loaded onto the instrument running a standard 30-minute gradient."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["Protein identification and quantification (RPL28 knockdown) A database search was performed using MaxQuant version 1.6.17 using label-free quantitation (LFQ).(Cox and Mann, 2008) Search parameters included the variable modifications methionine oxidation and N-terminal acetylation, and the fixed modification carbamidomethylation of cysteine residues. A 1% false discovery rate was applied at the protein and peptide level. Mass spectra were searched against a FASTA database for the Homo sapiens (H. sapiens) proteome from UniProt dated 2020.  Proteomic statistical and bioinformatic analyses (RPL28 knockdown) Bioinformatic analysis was performed in R using LFQ intensities for quantitation, and differential expression analysis was performed using limma. Protein identification and quantification (BOP1 knockdown) A database search was performed using DIA-NN version 1.8.1 using label-free quantitation (LFQ) (Cox and Mann, 2008). Mass spectra were searched against a FASTA database for the H. sapiens proteome from UniProt. Search parameters included the fixed modifications N-terminal methionine excision and carbamidomethylation of cysteine residues, with no variable modifications. A 1% false discovery rate was applied at the protein and peptide level. Proteomic statistical and bioinformatic analyses (BOP1 knockdown) Bioinformatic analysis was performed in RStudio using LFQ intensities for quantitation. Proteins were filtered to a minimum quantification of 25% across all samples. Filtered data was fitted to a linear model by limma (Ritchie et al., 2015). Gene ontology (GO) enrichment analyses were performed by Panther, which performed over-representation analyses by Fisher’s exact test and calculated false discovery rates accordingly (Mi et al., 2019; Thomas et al., 2022)."],"omics_type":["Proteomics"],"labhead":["Leonard J"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Department of Biochemistry and Molecular Biology; Michael Smith Laboratories; University of British Columbia"],"species":["Homo Sapiens (human)"],"publication":["Not available"],"submitter_mail":["b_hay@live.ca"],"submitter_affiliation":["UBC"],"submitter_country":["Canada"],"additional_accession":[]},"is_claimable":false,"name":"Ribosome biogenesis mediates the translational increase of non-optimal codon transcripts during IFN stimulation","description":"The interferon response is a signaling pathway unique to vertebrates that links the innate and adaptive immune responses. Interferons signal through a cascade of factors including the JAK-STAT pathway to induce the transcription of hundreds of interferon stimulated genes (ISGs). Although the main interferon signal transduction pathways and ISGs have been elucidated, translational regulation of ISG transcripts has not been fully investigated. Prior work demonstrated that ribosomal protein RPL28 negatively regulates a subset of ISGs; however, we find that this effect may be due to a reduction in overall ribosome availability. Multi-omics analysis of RNA-seq and LC-MS/MS data reveal proteins that are translationally up-regulated in IFN-β-stimulated cells depleted of ribosome biogeneis factor BOP1, including an enrichment of ISGs. Analysis of codon usage demonstrates a significant reduction in codon optimality for proteins that are translationally up-regulated during BOP1 knockdown and IFN-β stimulation. Using reporter constructs, we demonstrate that codon non-optimal reporters are translated more that codon-optimized reporters in BOP1-depleted IFN-β cells. We propose that ribosome biogenesis regulates translational fine-tuning of integral protein production to ensure optimal interferon responses.","dates":{"publication":"2026-04-30","submission":"2025-01-26"},"accession":"PXD060209","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:1131","NEWT:35795","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:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:1073882","NEWT:6819","NEWT:475174","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","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:3760","NEWT:1584","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","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:3349","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:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","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:3715","NEWT:2065263","NEWT:177437","NEWT:717966","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","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:9006","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:485","NEWT:56636","NEWT:2499525","NEWT:2853422","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:2836","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:321967","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:415978","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:41122","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:29488","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231006","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:542","NEWT:2903","NEWT:185579","NEWT:872227","NEWT:13076","NEWT:2587410","NEWT:207340","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:2126337","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:64657","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","NCB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