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A central regulator of this adaptability is the stringent response, mediated by the alarmones (p)ppGpp. Activation of the stringent response triggers a global physiological shift away from active metabolism toward stress management and cellular quiescence, a key transition to stationary phase. To date, few studies have investigated the protein-level processes under stringent response control. In this study, we performed shotgun proteomic profiling (LC-MS\/MS) of a (p)ppGpp-null\n                      <jats:italic toggle=\"yes\">relA spoT<\/jats:italic>\n                      double knockout mutant (\u0394SR) and its isogenic PAO1 parent during late exponential and stationary phase planktonic growth in rich medium. In this first characterization of the stringent response proteome in\n                      <jats:italic toggle=\"yes\">P.a<\/jats:italic>\n                      , we identified 146 differentially abundant proteins (DAP) in late exponential phase \u0394SR cells, and 2.3-fold greater DAP (339) in stationary-phase cells. Functional characterization and system-level analysis of the differentially abundant stationary phase proteome highlighted the key role of (p)ppGpp in shutting down a wide range of metabolic processes while inducing adaptive processes such as responses to oxidative stress. Comparative RNA-seq analysis also uncovered incongruent protein and mRNA responses in the \u0394SR relative to wild-type cells in several processes, including iron homeostasis and antioxidant defenses. Transcriptomic analysis of the \u0394SR also revealed differential expression of numerous sRNAs, including PrrF1\/2, raising the possibility that sRNAs may mediate certain stringent-response-dependent post-transcriptional control. Phenotypic validation of the \u0394SR proteome confirmed multiple defects in antioxidant defenses and dysregulated pyoverdine biosynthesis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Pseudomonas aeruginosa<\/jats:italic>\n                      is an important opportunistic human pathogen that causes difficult-to-treat infections and is highly adaptable to stressful environments. A key orchestrator of this adaptability is the stringent response, mediated by the signaling molecule (p)ppGpp. In this study, we characterized the extensive proteome remodeling in the (p)ppGpp-null\n                      <jats:italic toggle=\"yes\">relA spoT<\/jats:italic>\n                      mutant during late exponential and stationary phase and provided the first protein-level analysis of the stringent response in\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      . These data highlight the role of (p)ppGpp in shutting down broad metabolic functions and activating adaptive pathways, including stress-resistance responses, particularly during stationary phase. Using comparative proteomics and RNA-seq, we also find that several processes\u2014such as iron regulation and oxidative stress responses\u2014show mismatched RNA and protein changes, suggesting additional layers of post-transcriptional control, potentially involving small regulatory RNAs.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/msystems.01742-25","type":"journal-article","created":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T13:03:45Z","timestamp":1789650225000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Proteomic profiling of the\n                    <i>Pseudomonas aeruginosa<\/i>\n                    stringent response highlights global adaptive responses during stationary phase"],"prefix":"10.1128","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-8555-5581","authenticated-orcid":true,"given":"Jacquelyn","family":"Rich","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01pxwe438","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, McGill University","place":["Montreal, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6886-6933","authenticated-orcid":false,"given":"Samuel J. 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