biostudies-literatureLin YSNational Taiwan University HospitalMinistry of EducationNational Taiwan UniversityNational Health Research InstitutesAcademia SinicaNational Science and Technology CouncilKaohsiung Veterans General Hospitale20220727https://www.ebi.ac.uk/biostudies/studies/S-EPMC10174191biostudies-literatureUnknown220(8)Type I interferons are important antiviral cytokines, but prolonged interferon production is detrimental to the host. The TLR3-driven immune response is crucial for mammalian antiviral immunity, and its intracellular localization determines induction of type I interferons; however, the mechanism terminating TLR3 signaling remains obscure. Here, we show that the E3 ubiquitin ligase ZNRF1 controls TLR3 sorting into multivesicular bodies/lysosomes to terminate signaling and type I interferon production. Mechanistically, c-Src kinase activated by TLR3 engagement phosphorylates ZNRF1 at tyrosine 103, which mediates K63-linked ubiquitination of TLR3 at lysine 813 and promotes TLR3 lysosomal trafficking and degradation. ZNRF1-deficient mice and cells are resistant to infection by encephalomyocardThe Journal of experimental medicineThe Src-ZNRF1 axis controls TLR3 trafficking and interferon responses to limit lung barrier damage.PMC10174191109-2327-B-002-009NTU-CC-111L893903110-2634-F-002-044AS-CFII-108-104ASCFII-108-104111C101-7108-2320-B-002-020-MY3KSVGH110-025109-2314-B-002-080110L901402BNHRI-EX110-11031SI109-2320-B-002-018-MY3110-2740-B-002-006ASIA-106-L04Tsai CYHo YHTsai YMLin YSChao TLLai TYLee CYJhuang SJHsu LCChang SYHsueh YPChang YCChen CYChuang THLiu YLfalseThe Src-ZNRF1 axis controls TLR3 trafficking and interferon responses to limit lung barrier damage.Type I interferons are important antiviral cytokines, but prolonged interferon production is detrimental to the host. The TLR3-driven immune response is crucial for mammalian antiviral immunity, and its intracellular localization determines induction of type I interferons; however, the mechanism terminating TLR3 signaling remains obscure. Here, we show that the E3 ubiquitin ligase ZNRF1 controls TLR3 sorting into multivesicular bodies/lysosomes to terminate signaling and type I interferon production. Mechanistically, c-Src kinase activated by TLR3 engagement phosphorylates ZNRF1 at tyrosine 103, which mediates K63-linked ubiquitination of TLR3 at lysine 813 and promotes TLR3 lysosomal trafficking and degradation. ZNRF1-deficient mice and cells are resistant to infection by encephalomyocard2023-01-01T00:00:00Z2023 Aug2026-07-15T01:42:19.055Z2025-04-05T21:09:00.33ZS-EPMC101741913715898210.1084/jem.20220727