article published yes Murat Artan author C407B586-6052-11E9-B3AE-7006E66974250000-0001-8945-6992 Hanna Schön author C8E17EDC-D7AA-11E9-B7B7-45ECE5697425 Mario De Bono author 4E3FF80E-F248-11E8-B48F-1D18A9856A870000-0001-8347-0443 MaDe department ISTplus - Postdoctoral Fellowships project Molecular mechanisms of neural circuit function project Insulin/insulin-like growth factor signaling inhibits FOXO transcription factors to control development, homeostasis, and aging. Here, we use proximity labeling to identify proteins interacting with the C. elegans FOXO DAF-16. We show that in well-fed, unstressed animals harboring active insulin signaling, DAF-16 forms a complex with the PAR-1/MARK serine/threonine kinase, a key regulator of cell polarity. PAR-1 inhibits DAF-16 accumulation and promotes DAF-16 phosphorylation at S249, at a conserved motif that PAR-1/human MARK2 phosphorylates in vitro. DAF-2 insulin-like receptor signaling stimulates DAF-16 S249 phosphorylation, suggesting DAF-2 activates PAR-1. DAF-2 also promotes PAR-1 expression by inhibiting DAF-16. PAR-1 knockdown, or DAF-16 S249A, prolong lifespan, whereas phosphomimetic DAF-16 S249D suppresses the longevity of daf-2 mutants. At low insulin signaling, DAF-16 proximity labeling highlights transcription factors, chromatin regulators, and DNA repair proteins. One interactor, the zinc finger/homeobox protein ZFH-2/ZFHX3, forms a complex with DAF-16 and prolongs lifespan. Our work provides entry points for hypothesis-driven studies of FOXO function and longevity. https://research-explorer.ista.ac.at/download/20929/20941/2025_NatureComm_Artan.pdf application/pdfno Springer Nature2025 eng 2041-1723 4138145210.1038/s41467-025-66409-0 16 Artan M, Schön H, de Bono M. 2025. Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins. Nature Communications. 16, 11355. Artan, M., Schön, H., &#38; de Bono, M. (2025). Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins. <i>Nature Communications</i>. Springer Nature. <a href="https://doi.org/10.1038/s41467-025-66409-0">https://doi.org/10.1038/s41467-025-66409-0</a> M. Artan, H. Schön, M. de Bono, Nature Communications 16 (2025). Artan, Murat, et al. “Proximity Labeling of DAF-16 FOXO Highlights Aging Regulatory Proteins.” <i>Nature Communications</i>, vol. 16, 11355, Springer Nature, 2025, doi:<a href="https://doi.org/10.1038/s41467-025-66409-0">10.1038/s41467-025-66409-0</a>. Artan, Murat, Hanna Schön, and Mario de Bono. “Proximity Labeling of DAF-16 FOXO Highlights Aging Regulatory Proteins.” <i>Nature Communications</i>. Springer Nature, 2025. <a href="https://doi.org/10.1038/s41467-025-66409-0">https://doi.org/10.1038/s41467-025-66409-0</a>. M. Artan, H. Schön, and M. de Bono, “Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins,” <i>Nature Communications</i>, vol. 16. Springer Nature, 2025. Artan M, Schön H, de Bono M. Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins. <i>Nature Communications</i>. 2025;16. doi:<a href="https://doi.org/10.1038/s41467-025-66409-0">10.1038/s41467-025-66409-0</a> 209292026-01-04T23:01:34Z2026-05-20T08:10:18Z