Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins

Artan, Murat; Schön, Hanna; De Bono, Mario

Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins

Artan M, Schön H, de Bono M. 2025. Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins. Nature Communications. 16, 11355.

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DOI

10.1038/s41467-025-66409-0

Journal Article | Published | English

Scopus indexed

Author

Artan, Murat^ISTA ; Schön, Hanna^ISTA; de Bono, Mario^ISTA

Corresponding author has ISTA affiliation

Department

De Bono Group

Grant

ISTplus - Postdoctoral Fellowships
Molecular mechanisms of neural circuit function

Abstract

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.

Publishing Year

2025

Date Published

2025-12-11

Journal Title

Nature Communications

Publisher

Springer Nature

Acknowledgement

We thank de Bono lab members for helpful comments on the manuscript, and the Mass Spec Facility at the Max Perutz Labs, notably WeiQiang Chen and Markus Hartl, for invaluable discussions and comments on mass spec analyses of worm samples. All LC-MS/MS analyses were performed on instruments of the Vienna BioCenter Core Facilities (VBCF). Microscopy was supported by the Scientific Services Units (SSU) of ISTA through resources provided by the Imaging & Optics Facility (IOF). We are grateful to Dr. Geraldine Seydoux (Johns Hopkins University) for worm strains and plasmids, and Dr. Seung-Jae V. Lee (KAIST) for RNAi clones. We are grateful to Ekaterina Lashmanova for designing the daf-16::TbID::mNG::3xFLAG knock-in construct and for her outstanding support in the lab. This work was supported by a Wellcome Investigator Award (209504/A/17/Z) to MdB and an ISTplus Fellowship to MA (Marie Sklodowska-Curie agreement No 754411).

Acknowledged SSUs

Imaging & Optics Facility

Volume

Article Number

11355

eISSN

2041-1723

IST-REx-ID

20929

Cite this

Artan M, Schön H, de Bono M. Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins. Nature Communications. 2025;16. doi:10.1038/s41467-025-66409-0

Artan, M., Schön, H., & de Bono, M. (2025). Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins. Nature Communications. Springer Nature. https://doi.org/10.1038/s41467-025-66409-0

Artan, Murat, Hanna Schön, and Mario de Bono. “Proximity Labeling of DAF-16 FOXO Highlights Aging Regulatory Proteins.” Nature Communications. Springer Nature, 2025. https://doi.org/10.1038/s41467-025-66409-0.

M. Artan, H. Schön, and M. de Bono, “Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins,” Nature Communications, vol. 16. Springer Nature, 2025.

Artan M, Schön H, de Bono M. 2025. Proximity labeling of DAF-16 FOXO highlights aging regulatory proteins. Nature Communications. 16, 11355.

Artan, Murat, et al. “Proximity Labeling of DAF-16 FOXO Highlights Aging Regulatory Proteins.” Nature Communications, vol. 16, 11355, Springer Nature, 2025, doi:10.1038/s41467-025-66409-0.

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