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bioRxiv · 10.64898/2025.12.29.696917

PtdIns(3,5)P2 Is an Endogenous Ligand of STING in Innate Immune Signaling

Tan, J. X.·Lv, B.·Li, J.·Li, T.·Du, F.·Chen, X.·Zhang, X.·Bai, X.-c.·Chen, Z. J.
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Abstract

Cytosolic DNA exposure triggers innate immune responses through cyclic GMP-AMP (cGAMP) synthase (cGAS)1-3. Upon binding to DNA, cGAS is activated to produce cGAMP, which functions as a second messenger that binds to stimulator of interferon genes (STING), an endoplasmic reticulum (ER)-localized signaling adaptor3-5. STING then traffics from the ER to the Golgi, leading to activation of the kinases TBK1 and IKK and subsequent induction of interferons and other cytokines6-10. Here we show that phosphatidylinositol 3,5-bisphosphate [PtdIns(3,5)P2] is an endogenous ligand of STING that functions together with cGAMP to induce STING activation. Proteomics analysis identified a constitutive interaction between STING and PIKfyve, an enzyme that produces PtdIns(3,5)P2 in mammalian cells. Deletion of PIKfyve blocked STING trafficking from the ER and TBK1 activation. In vitro reconstitution revealed a strong and selective effect of PtdIns(3,5)P2 on STING activation. Purified STING bound directly to PtdIns(3,5)P2 in a fluorescence resonance energy transfer (FRET) assay. Consistently, PtdIns(3,5)P2 promoted cGAMP-induced STING oligomerization by binding to a groove between STING dimers as revealed by cryo-EM (Li et al., co-submitted). Similar to PIKfyve depletion, mutation of the PtdIns(3,5)P2-binding residues in STING blocked its trafficking and downstream signaling. These results reveal PtdIns(3,5)P2 as a lipid ligand of STING with essential roles in innate immunity.

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Tan, J. X., Lv, B., Li, J., Li, T., Du, F., Chen, X., Zhang, X., Bai, X.-c., Chen, Z. J.. 2025-12-29. PtdIns(3,5)P2 Is an Endogenous Ligand of STING in Innate Immune Signaling. https://doi.org/10.64898/2025.12.29.696917

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