A Spatiotemporal Atlas of Extranuclear Androgen Receptor Proximal Interaction Networks

Celeste C Ptak; Conor O'Rourke; Jimmy K Eng; Lilliana Radoshevich; Michael E Wright

doi:10.64898/2026.08.03.742469

Androgen receptor-interacting proteins (AR-IPs) comprise nearly 1,000 proteins, yet their organization across subcellular space and time remains uncharted. Proximity labeling captures direct binding partners along with neighboring proteins that populate a receptor's local environment, thereby broadening AR-IPs into a broader population of AR-proximal interacting proteins (AR-PIPs). Here, we apply proximity labeling quantitative mass spectrometry (PL-qMS) to construct a spatiotemporal atlas of the extranuclear AR-proximal interactome in LNCaP prostate tumor cells. PL-qMS recovered 82.2% of the known AR-IPs and identified 3,947 AR-PIPs across cytosolic and membrane compartments, revealing dynamic remodeling across an androgen time course. Functional enrichment and network analyses identified the retromer complex as an androgen-sensitive AR-proximal interaction, which was verified by proximity ligation assays. Partial genetic disruption of VPS26A attenuated transcription of canonical androgen-regulated genes by mislocalizing the AR coactivator TMF1, establishing the retromer-AR-TMF1 axis as a functionally validated AR-proximal interaction network (AR-PIN). This work establishes subcellular proximal proteomes as a spatiotemporal framework for probing AR function and its dysregulation in disease.Androgen receptor-interacting proteins (AR-IPs) comprise nearly 1,000 proteins, yet their organization across subcellular space and time remains uncharted. Proximity labeling captures direct binding partners along with neighboring proteins that populate a receptor's local environment, thereby broadening AR-IPs into a broader population of AR-proximal interacting proteins (AR-PIPs). Here, we apply proximity labeling quantitative mass spectrometry (PL-qMS) to construct a spatiotemporal atlas of the extranuclear AR-proximal interactome in LNCaP prostate tumor cells. PL-qMS recovered 82.2% of the known AR-IPs and identified 3,947 AR-PIPs across cytosolic and membrane compartments, revealing dynamic remodeling across an androgen time course. Functional enrichment and network analyses identified the retromer complex as an androgen-sensitive AR-proximal interaction, which was verified by proximity ligation assays. Partial genetic disruption of VPS26A attenuated transcription of canonical androgen-regulated genes by mislocalizing the AR coactivator TMF1, establishing the retromer-AR-TMF1 axis as a functionally validated AR-proximal interaction network (AR-PIN). This work establishes subcellular proximal proteomes as a spatiotemporal framework for probing AR function and its dysregulation in disease.Proximity labeling constructs a spatiotemporal atlas of the extranuclear AR-proximal interactome and identifies the retromer complex as an androgen-sensitive regulator of AR transcription. PL-qMS constructs a spatiotemporal atlas of the extranuclear AR-proximal interactomeAR-PIPs recover 3,947 proximal interactors across cytosolic and membrane fractionsRetromer complex is an androgen-sensitive AR-proximal interactionPartial VPS26A disruption attenuates AR-dependent gene transcription via TMF1 mislocalization.SynopsisProximity labeling constructs a spatiotemporal atlas of the extranuclear AR-proximal interactome and identifies the retromer complex as an androgen-sensitive regulator of AR transcription. PL-qMS constructs a spatiotemporal atlas of the extranuclear AR-proximal interactomeAR-PIPs recover 3,947 proximal interactors across cytosolic and membrane fractionsRetromer complex is an androgen-sensitive AR-proximal interactionPartial VPS26A disruption attenuates AR-dependent gene transcription via TMF1 mislocalization.

A Spatiotemporal Atlas of Extranuclear Androgen Receptor Proximal Interaction Networks

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