TBC1D23-AAVR interaction drives endosome-to-TGN trafficking required for rAAV transduction

Xiujuan Zhang; Ariful Habib; Shane McFarlin; Donovan Richart; Soo Yeun Park; Fang Cheng; Jinxi Wang; Ziying Yan; Jianming Qiu

doi:10.1073/pnas.2605086123

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TBC1D23-AAVR interaction drives endosome-to-TGN trafficking required for rAAV transduction

Journal article

Open access

Peer reviewed

TBC1D23-AAVR interaction drives endosome-to-TGN trafficking required for rAAV transduction

Xiujuan Zhang, Ariful Habib, Shane McFarlin, Donovan Richart, Soo Yeun Park, Fang Cheng, Jinxi Wang, Ziying Yan and Jianming Qiu

Proceedings of the National Academy of Sciences - PNAS, Vol.123(33), e2605086123

08/18/2026

DOI: 10.1073/pnas.2605086123

PMID: 42584998

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Abstract

The adeno-associated virus receptor (AAVR; also known as KIAA0319L) is the proteinaceous receptor required for the transduction of multiserotype adeno-associated viruses (AAVs). While the extracellular polycystic kidney disease (PKD) domains of AAVR directly bind AAV capsids, the function of its C-terminal cytosolic domain (AAVR-C) in governing AAV internalization and intracellular trafficking remains undefined. Using targeted pulldown of AAVR-interacting host proteins with a glutathione S-Transferase (GST)-fused AAVR-C recombinant protein (GST-AAVR-C) as a bait, we identified that TBC1 domain family member 23 (TBC1D23), a specialized intracellular trafficking adaptor and bridging factor, binds AAVR via the AAVR-C and mediates AAV's endosome-to- -Golgi network (TGN) transport. Biolayer interferometry (BLI) demonstrates nanomolar-affinity binding between AAVR-C and the C-terminal scaffold domain of TBC1D23 (TBC1D23-C). CRISPR-mediated knockout of severely impairs rAAV transduction across multiple human cell types, including polarized human airway epithelium (HAE). Loss of TBC1D23 disrupts convergence of internalized AAV capsids at the TGN, leading to diminished nuclear import of AAV vectors. Mutational analyses of AAVR-C reveal that its acidic residue cluster is required for TBC1D23 binding and AAV retrograde transport from the endosome to the TGN. Together, these findings define TBC1D23 as a receptor-proximal trafficking module that couples AAV-AAVR engagement to vesicle transport, revealing a missing core regulatory step in AAV retrograde transport that is essential for productive rAAV transduction.

Dependovirus - genetics

Dependovirus - metabolism

Endosomes - metabolism

GTPase-Activating Proteins - genetics

GTPase-Activating Proteins - metabolism

HEK293 Cells

Humans

Protein Binding

Protein Transport

Receptors, Cell Surface

trans-Golgi Network - metabolism

Transduction, Genetic

Details

Title: Subtitle: TBC1D23-AAVR interaction drives endosome-to-TGN trafficking required for rAAV transduction
Creators: Xiujuan Zhang - University of Kansas Medical Center
Ariful Habib - University of Kansas Medical Center
Shane McFarlin - University of Kansas Medical Center
Donovan Richart - University of Kansas Medical Center
Soo Yeun Park - University of Alabama at Birmingham
Fang Cheng - University of Kansas Medical Center
Jinxi Wang - University of Kansas Medical Center
Ziying Yan - University of Alabama at Birmingham
Jianming Qiu - University of Kansas Medical Center
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.123(33), e2605086123
DOI: 10.1073/pnas.2605086123
PMID: 42584998
ISSN: 1091-6490
eISSN: 1091-6490
Grant note: HL174593 / HHS | NIH (NIH) AI180416 / HHS | NIH (NIH) AI182645 / HHS | NIH (NIH) YAN23G0 / Cystic Fibrosis Foundation (CFF) AI166293 / HHS | NIH (NIH)
Language: English
Date published: 08/18/2026
Academic Unit: Anatomy and Cell Biology
Record Identifier: 9985218398602771

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