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Weldemariam"],"technology_type":["Mass Spectrometry","Bottom-up proteomics"],"software":[""],"submitter_keywords":["Phus","Prrf small rnas","Heme-binding protein","Pseudomonas aeruginosa","H209a variant"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD079786"],"sample_protocol":["Pseudomonas aeruginosa static samples were harvested by scraping using a sterile loop and were frozen at -80°C until ready for processing Protein extraction and digestion were performed using S-trap micro columns (ProtiFi, NY) according to the manufacturer’s instructions as previously described.  Briefly, samples were lysed in a lysis buffer containing 5% sodium dodecyl sulfate (Sigma, L4509), 50 mM triethylammonium bicarbonate (1 M, pH 8.0) (Sigma, 7408). Proteins were extracted and digested using S-trap micro columns (ProtiFi, NY). The eluted peptides from the S-trap column were dried, and peptide concentration was determined using a BCA assay kit (Thermo Fisher Scientific, 23275), after reconstitution in 0.1% formic acid. LC-MS/MS-based proteomic analysis was conducted on a nanoACQUITY Ultra-Performance Liquid Chromatography system (Waters Corporation, Milford, MA USA) coupled to an Orbitrap Fusion Lumos Tribrid mass spectrometer (Thermo Scientific, San Jose, CA USA). Peptide separation was affected on a nanoACQUITY Ultra-Performance Liquid Chromatography (UPLC) analytical column (BEH130 C18, 1.7 µm, 75 µm x 200 mm; Waters Corporation, Milford, MA, USA) using a 185-min linear gradient with 3-40% acetonitrile and 0.1% formic acid."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["The resulting mass spectra were processed using Thermo Proteome Discoverer ((PD, version 2.5.0.400), Thermo Fisher Scientific) and searched against a UniProt Pseudomonas Aeruginosa reference proteome (release 2024.03, 1526 entries) using Sequest HT algorithm. Search parameters include carbamidomethylation of cysteines (+57.021 Da) as a static modification, methionine oxidation (+15.995 Da) as a dynamic modification, precursor mass tolerance of 20 ppm, fragment mass tolerance of 0.5 Da, and trypsin as a digestion enzyme. Tryptic missed cleavages were restricted to a maximum of two, with peptide lengths set between 6 and 144 residues. For protein quantification, the Minora feature detector, integrated in the PD, was used as previously described (73). To ensure high data quality, proteins were further filtered to a 1% false discovery rate (FDR) threshold, calculated with the Percolator algorithm. Next, protein abundance values exported from PD were post-processed using Perseus software (version 1.6.14.0). Proteins with missing values were excluded to improve data quality. The quantitative protein data were log2 transformed and further normalized using median centering. Differentially expressed proteins (DEPs) were identified using a two-tailed Student’s t-test (adjusted p-value < 0.05). Ingenuity Pathway Analysis (IPA) software (Qiagen, Germantown, MD) was used to identify dysregulated pathways and biological processes."],"omics_type":["Proteomics"],"labhead":["Maureen A Kane"],"instrument_platform":[""],"labhead_affiliation":["Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland Baltimore"],"submission_type":["PARTIAL"],"species":["Pseudomonas Aeruginosa Pa103"],"publication":["Not available"],"submitter_mail":["mweldemariam@rx.umaryland.edu"],"submitter_affiliation":["University of Maryland Baltimore"],"submitter_country":["United States"],"additional_accession":[]},"is_claimable":false,"name":"Disruption of Heme Uptake and PrrF sRNA Expression in the Pseudomonas aeruginosa PhuS H209A Variant Induces a Metabolic Stress-Response","description":"The Pseudomonas aeruginosa heme-binding protein PhuS plays a central role in coordinating heme utilization with iron homeostasis through the PrrF small RNAs. Proteomic and metabolomic analyses of the phuSH209A allelic strain demonstrated that the previously characterized in vitro disruption of PhuS function results in widespread regulatory and metabolic perturbations, through its influence on the differential expression of the PrrF small RNAs. Although intracellular iron levels remain unchanged relative to PAO1, the phuSH209A allelic strain exhibits decreased expression of pyochelin (PCH) biosynthetic proteins and increased abundance of iron uptake receptors. This perceived iron dysregulation is consistent with  disruption of the PrrF-dependent iron-sparing response, suggesting that PhuS functions beyond heme trafficking as a modulator of iron-dependent post-transcriptional regulation. Specifically, the differential expression of PrrF1 and PrrF2 under heme conditions indicates that PhuS impacts the balance and activity of these sRNAs, which govern iron-responsive metabolic pathways. This includes decreased conservation of anthranilate for quorum sensing (QS) signaling molecules, contributing to reduced Pseudomonas quinolone signal (PQS) production and downstream effects on phenazine and siderophore biosynthesis.","dates":{"publication":"2026-09-28","submission":"2026-06-17"},"accession":"PXD079786","cross_references":{"TAXONOMY":["NEWT:3555","NEWT:71647","NEWT:330879","NEWT:35554","NCBITaxon:12939","NEWT:32046","NEWT:544496","NEWT:2042546","NEWT:32049","NEWT:1392696","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:45351","NEWT:445974","NEWT:43179","NEWT:180454","NEWT:5722","NCBITaxon:1280","NEWT:1129","NEWT:309807","NEWT:10245","NEWT:55153","NEWT:10244","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:351581","NEWT:43186","NEWT:428146","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:498019","NEWT:1351","NEWT:1438992","NEWT:1352","NEWT:2649997","NEWT:1147161","NEWT:661410","NEWT:638632","NEWT:263737","NEWT:224326","NEWT:1333499","NCBITaxon:79857","NEWT:1096976","NEWT:5702","NEWT:95648","NEWT:1589","NEWT:135622","NEWT:1349","NEWT:67352","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:383379","NEWT:1401257","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:44491","NEWT:641809","NEWT:27933","NEWT:556484","NEWT:317447","NEWT:4688","NEWT:7955","NEWT:7719","NEWT:7959","NEWT:2261","NEWT:868565","NEWT:31156","NEWT:3112","NEWT:79329","NEWT:4442","NEWT:31155","NEWT:192875","NEWT:1214915","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:1316931","NEWT:224308","NEWT:3347","NEWT:511145","NEWT:160621","NEWT:212790","NEWT:931281","NEWT:4432","NEWT:5762","NEWT:5763","NEWT:658457","NEWT:1310161","NEWT:77133","NEWT:295358","NEWT:145481","NCBITaxon:79824","NEWT:2246","NEWT:2652724","NEWT:1912919","NEWT:5757","NCBITaxon:4563","NEWT:5755","NEWT:44689","NEWT:44447","NEWT:752555","NEWT:498211","NEWT:5759","NEWT:347256","NEWT:1736231","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:1392","NEWT:498217","NEWT:498216","NEWT:2242","NEWT:11320","NEWT:286","NEWT:391619","NEWT:246196","NEWT:287","NEWT:246197","NEWT:1367269","NEWT:145458","NEWT:633149","NEWT:1149786","NEWT:10239","NEWT:68895","NEWT:44685","NEWT:161934","NEWT:4897","NEWT:1148","NEWT:5744","NEWT:5508","NEWT:3329","NEWT:5507","NEWT:410661","NEWT:55571","NEWT:35500","NEWT:1235816","NEWT:1140","NCBITaxon:2157","NEWT:1143","NEWT:4896","NEWT:1287689","NEWT:1390","NEWT:11557","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:294","NEWT:1773","NEWT:1182590","NEWT:3712","NEWT:82380","NEWT:3711","NEWT:935293","NEWT:375146","NEWT:270643","NEWT:2065263","NEWT:1772","NEWT:118698","NEWT:1616117","NEWT:263","NEWT:118696","NEWT:52283","NEWT:11988","NEWT:284812","NEWT:8175","NEWT:400667","NEWT:43330","NEWT:980415","NEWT:1603293","NEWT:408169","NEWT:44664","NEWT:47946","NEWT:459758","NEWT:3702","NEWT:1245466","NEWT:243277","NEWT:1246791","NEWT:7067","NEWT:118694","NEWT:586722","NEWT:2850","NEWT:118691","NEWT:33548","NEWT:274","NEWT:7070","NCBITaxon:287","NEWT:408172","NEWT:408170","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1436733","NEWT:460519","NEWT:1247411","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:9483","NEWT:333760","NEWT:56636","NEWT:1513458","NEWT:1233681","NCBITaxon:40559","NEWT:506599","NEWT:2853422","NEWT:81077","NEWT:84588","NEWT:1679718","NEWT:480","NEWT:65349","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:300852","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:508771","NEWT:1883446","NEWT:253","NCBITaxon:2759","NEWT:1233435","NEWT:93061","NEWT:93062","NEWT:34613","NCBITaxon:4932","NEWT:109760","NEWT:943274","NEWT:644223","NEWT:235443","NEWT:108458","NEWT:5936","NEWT:272623","NEWT:272624","NEWT:320637","NEWT:2719099","NEWT:886559","NEWT:55521","NEWT:983964","NEWT:118499","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:1130798","NEWT:55529","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:476272","NEWT:3747","NEWT:195051","NEWT:3988","NEWT:1116234","NEWT:1255228","NEWT:649908","NEWT:410289","NEWT:373153","NEWT:3983","NEWT:203696","NEWT:352472","NEWT:1071661","NEWT:1202532","NEWT:360094","NEWT:470","NEWT:41364","NEWT:5911","NEWT:1313","NEWT:1218097","NCBITaxon:50557","NEWT:39251","NEWT:39491","NEWT:212142","NCBITaxon:5811","NEWT:29491","NEWT:101841","NEWT:2014887","NEWT:33952","NEWT:153009","NEWT:261756","NEWT:571256","NEWT:40483","NEWT:63366","NEWT:63367","NEWT:688333","NEWT:215402","NCBITaxon:548681","NEWT:318586","NCBITaxon:5820","NEWT:1547","NEWT:272634","NEWT:198107","NEWT:9031","NEWT:1872122","NEWT:1308","NEWT:7091","NEWT:1034015","NEWT:760568","NEWT:141262","NEWT:108931","NEWT:1055524","NEWT:4087","NEWT:9778","NEWT:306","NEWT:150475","NEWT:303","NEWT:267872","NEWT:97477","NEWT:7111","NEWT:347515","NEWT:172269","NEWT:9534","NEWT:6269","NEWT:5180","NEWT:256737","NEWT:4090","NEWT:4092","NEWT:9541","NEWT:8694","NEWT:4093","NEWT:4096","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:941442","NEWT:536088","NEWT:13076","NEWT:1226408","NEWT:7108","NEWT:40479","NEWT:43989","NEWT:4076","NEWT:142615",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