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Whereas activation of STATs by JAKs via canonical receptor-driven JAK\u2013STAT signaling is well understood, little is known about JAK-independent mechanisms of STAT activation. Here, we identify the understudied nonreceptor tyrosine kinase TNK1 as a therapeutically targetable, JAK-independent activator of STAT signaling. Using a multiomics approach, we mapped a network of TNK1 substrates associated with protein condensates and proinflammatory signaling, including STAT1. We found that TNK1, but not its sister kinase ACK1, directly phosphorylates STATs at well described STAT-activating JAK sites. In cells, TNK1-mediated STAT1 phosphorylation and activation occurs independently of JAKs. Imaging and interactomics data suggest that TNK1 interacts with STAT1 in cytosolic condensates, which likely compartmentalize TNK1-substrate interactions. We show that an intrinsically disordered proline-rich region in TNK1, which includes a 14-3-3 docking phosphorylation site, is required for the formation of kinase-active TNK1 condensates and STAT1 phosphorylation. Mutations within the proline-rich region that eliminate 14-3-3 binding increase formation of TNK1 condensates, suggesting a model in which 14-3-3 acts as a clamp that constrains the flexible PRR to inhibit condensate formation and STAT1 activation. Finally, we show that TNK1 is a targetable driver of STAT1-mediated inflammation in the gut as inhibition of TNK1 reduces active STAT1 in the colon and ameliorates colitis symptoms in mice.<\/jats:p>","DOI":"10.1101\/gad.353368.125","type":"journal-article","created":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T17:00:24Z","timestamp":1785776424000},"page":"1408-1428","source":"Crossref","is-referenced-by-count":0,"title":["TNK1 is a targetable JAK-independent driver of STAT signaling and inflammation"],"prefix":"10.1101","volume":"40","author":[{"given":"Tania P.","family":"L\u00f3pez-Palacios","sequence":"first","affiliation":[{"name":"Department of Oncological Sciences, University of Utah, Salt Lake City, Utah 84112, USA;"},{"name":"Fritz B. 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