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Contrary to this paradigm, rapid nuclear translocation of phosphorylated STAT1 (pSTAT1) following cytokine stimulation requires importin \u03b2, Ran\u2013GTP, and the importin \u03b15 isoform, which recognizes a non-transferable NLS. Here, we present cryo-EM structures of pSTAT1 bound to importin \u03b15, revealing an asymmetric 2:1 complex that diverges from canonical NLS-mediated cargo recognition. Importin \u03b15 occupies the DNA-binding groove of the pSTAT1 dimer, with a single STAT1 N-terminal domain positioning the C-terminal Armadillo repeats 9\u201310 (S1B domain) orthogonal to the DNA-binding interface. This interface is also targeted by the Ebola virus protein VP24, an antagonist of interferon signaling. We further show that Ran\u2013GTP alone is insufficient to trigger nuclear release of pSTAT1, which additionally requires the exportin CAS. A cryo-EM reconstruction of the CAS\u2013Ran\u2013GTP\u2013\u03b15 complex, supported by in vitro competition assays, demonstrates that CAS and pSTAT1 are mutually exclusive ligands for importin \u03b15. Together, these findings define the molecular choreography of cytokine-induced STAT1 nuclear translocation and release, establishing a general paradigm for STAT family signaling.<\/jats:p>","DOI":"10.64898\/2026.09.28.754937","type":"posted-content","created":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T15:25:21Z","timestamp":1790695521000},"source":"Crossref","is-referenced-by-count":0,"title":["Cytokine-induced nuclear translocation of STAT1 via a non-transferable NLS"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2094-6683","authenticated-orcid":false,"given":"Pramod","family":"Kumar","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4515-0515","authenticated-orcid":false,"given":"James 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