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The PilY1 protein is a component of the type IV pili (T4P) but has additional signaling functions outside of the pilus. A major function of PilY1 is to prevent the AlgZ\/R two-component system from functioning. However, the complete effects of PilY1 signaling through the control of the AlgZ\/R or other regulatory systems are not well understood. In this study, we determine new genes controlled by the AlgZ\/R system that are impacted by PilY1. We discovered that PilY1 impacts cAMP by preventing the AlgZ\/R system from activating the major adenylate cyclase,\n                      <jats:italic toggle=\"yes\">cyaB<\/jats:italic>\n                      . PilY1 also prevents the AlgZ\/R system from activating itself and a putative c-di-GMP phosphodiesterase,\n                      <jats:italic toggle=\"yes\">PA4781<\/jats:italic>\n                      . Furthermore, PilY1 functions in different\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      strains, including mucoid strains. Surprisingly, bacterial survival in the lung, liver, and blood did not require PilY1. Overall, these findings suggest that PilY1 is critical for preventing AlgZ\/R activity to prevent improper second messenger production. This work identifies new AlgZ\/R targets and implicates the AlgZ\/R system in the control of cAMP in\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      . This work identifies a possible way to use PilY1 to prevent the expression of the major adenylate cyclase using PilY1 which could be used to impact\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      virulence.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Understanding PilY1 signaling is important for elucidating how\n                        <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                        adapts to different environments. A major function for PilY1 is to interfere with the AlgZ\/R two-component system. Here, we use transcriptomics to determine genes PilY1 affects through the AlgZ\/R system. We identified new AlgZ\/R targets and established a mechanism for impacting cAMP levels. Our findings further our understanding of PilY1 and the AlgZ\/R system and suggest a possible way that PilY1 might be used to prevent cAMP from increasing in\n                        <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                        .\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jb.00200-25","type":"journal-article","created":{"date-parts":[[2026,2,9]],"date-time":"2026-02-09T13:00:15Z","timestamp":1770642015000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Defining the role of\n                    <i>Pseudomonas aeruginosa<\/i>\n                    PilY1 in signaling and virulence"],"prefix":"10.1128","volume":"208","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2304-6680","authenticated-orcid":true,"given":"Christopher L.","family":"Pritchett","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01fpczx89","id-type":"ROR","asserted-by":"publisher"}],"name":"College of Public Health, East Tennessee State University","place":["Johnson City, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3140-402X","authenticated-orcid":true,"given":"F. 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