{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:04:24Z","timestamp":1785423864154,"version":"3.56.0"},"posted":{"date-parts":[[2026,1,6]]},"group-title":"Microbiology","reference-count":44,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,1,6]],"date-time":"2026-01-06T00:00:00Z","timestamp":1767657600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["R01AI099122"],"award-info":[{"award-number":["R01AI099122"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["Novel Therapeutics Development Grant"],"award-info":[{"award-number":["Novel Therapeutics Development Grant"]}],"id":[{"id":"https:\/\/ror.org\/033km2469","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,1,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  In this study, we investigated the impact of G protein-coupled receptor (GPCR) signaling on the intracellular replication of the model pathogen\n                  <jats:italic>Brucella neotomae<\/jats:italic>\n                  . Building on a prior chemical genetics screen, we identified agonists of the G\u03b1i-coupled adenosine A1 and dopamine D4 receptors as potent inhibitors of intracellular Brucella replication. In contrast, agonists of G\u03b1s-coupled adenosine A2A or dopamine D1 receptors, as well as antagonists of A1 or D4 receptors, either failed to inhibit or enhanced intracellular replication. Wild-type\n                  <jats:italic>B. neotomae<\/jats:italic>\n                  induced a rapid, type IV secretion system-dependent increase in host-cell cAMP during early infection. ENBA and cilostamide prevented this infection-associated cAMP increase and completely inhibited intracellular growth; this effect was partially reversed by cell-permeable cAMP analogs Using a real-time NanoBRET biosensor, we detected rapid G\u03b1s activation within minutes of infection that was sustained during wild-type but not \u0394virB4 infection and was abrogated by ENBA or cilostamide. Disruption of early G\u03b1s-cAMP signaling redirected BCVs to replication-incompatible phagolysosomal and autophagy-associated compartments. Collectively, these data support a model in which early GPCR signaling dynamics, balancing G\u03b1s and G\u03b1i pathways, are critical for establishment of productive intracellular Brucella infection.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Importance Statement<\/jats:title>\n                  <jats:p>Brucella species cause chronic infections by surviving and multiplying inside immune cells. To do this, Brucella must remodel the membrane-bound compartment that surrounds it after uptake, steering it away from destructive lysosomes and toward a permissive niche where replication can occur. We found that Brucella rapidly triggers a host signaling response controlled by G protein-coupled receptors, leading to a rise in a common cellular messenger molecule (cAMP) within minutes of infection. This early signal depends on the bacterial type IV secretion system and is required to build the replication-permissive compartment. When we disrupted this signaling with small molecules, bacteria were rerouted into degradative, autophagy-associated compartments and failed to establish productive infection. These results reveal an early host signaling checkpoint that Brucella exploits to create its intracellular niche and suggest that targeting host signaling dynamics, rather than bacterial viability directly, may provide new strategies to block intracellular infection.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.01.06.697936","type":"posted-content","created":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T03:05:10Z","timestamp":1767755110000},"source":"Crossref","is-referenced-by-count":1,"title":["Host GPCR-cAMP signaling balances G\u03b1s and G\u03b1i activity to control intracellular\n                  <i>Brucella<\/i>\n                  infection"],"prefix":"10.64898","author":[{"given":"Yoon-Suk","family":"Kang","sequence":"first","affiliation":[{"name":"Department of Pathology, Beth Israel Deaconess Medical Center & Harvard Medical School, Boston, MA, 02215, United States of 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