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Adeno-associated virus 2 bound to its cellular receptor AAVR

Nature Microbiology volume 4, pages 675–682 (2019)Cite this article

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Abstract

Adeno-associated virus (AAV) is a leading vector for virus-based gene therapy. The receptor for AAV (AAVR; also named KIAA0319L) was recently identified, and the precise characterization of AAV–AAVR recognition is in immediate demand. Taking advantage of a particle-filtering algorithm, we report here the cryo-electron microscopy structure of the AAV2–AAVR complex at 2.8 Å resolution. This structure reveals that of the five Ig-like polycystic kidney disease (PKD) domains in AAVR, PKD2 binds directly to the spike region of the AAV2 capsid adjacent to the icosahedral three-fold axis. Residues in strands B and E, and the BC loop of AAVR PKD2 interact directly with the AAV2 capsid. The interacting residues in the AAV2 capsid are mainly in AAV-featured variable regions. Mutagenesis of the amino acids at the AAV2–AAVR interface reduces binding activity and viral infectivity. Our findings provide insights into the biology of AAV entry with high-resolution details, providing opportunities for the development of new AAV vectors for gene therapy.

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Fig. 1: The cryo-EM structures.
Fig. 2: The complex structure of AAVR bound to AAV2.
Fig. 3: Conformational changes in the AAV2 capsid following AAVR attachment.
Fig. 4: Mutagenesis study of AAVR for AAV2 binding assays.

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Data availability

The obtained cryo-EM density maps and the data of the resolved structures were deposited into the Electron Microscopy Data Bank (EMDB) and Protein Data Bank (PDB) with the following accession numbers: AAV2, EMD-9671 and 6IH9; AAV2–AAVR complex, EMD-9672 and 6IHB. All other data supporting the findings of this study are available from the corresponding authors upon request.

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Acknowledgements

The authors thank the Computing and Cryo-EM Platforms of Tsinghua University, Branch of the National Center for Protein Sciences (Beijing) for providing facilities. They also thank L. Chen for his help in data collection, D. Yan for his assistance in molecular cloning, W. Huang for her help in cell culture, and Q. Ding for his discussion and comments. This work was supported by the National Program on Key Research Project of China (2017YFC0840300 and 2018YFA0507200) and the National Natural Science Foundation of China (grant numbers 81322023, 31770309, 81372284, 81520108019 and 31370733).

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Author notes

  1. These authors contributed equally: Ran Zhang, Lin Cao, Mengtian Cui.

Authors and Affiliations

  1. MOE Laboratory of Protein Science and Collaborative Innovation Center of Biotherapy, School of Medicine, Tsinghua University, Beijing, China

    Ran Zhang, Lin Cao, Zhiyong Lou & Zihe Rao

  2. School of Life Sciences, Tsinghua University, Beijing, China

    Ran Zhang, Zixian Sun, Mingxu Hu, Xueming Li & Zihe Rao

  3. State Key Laboratory of Medicinal Chemical Biology, College of Life Science and Pharmacy, Nankai University, Tianjin, China

    Lin Cao, Rouxuan Zhang & Zihe Rao

  4. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Capital Medical University, Beijing, China

    Mengtian Cui, Shentao Li & Wei Ding

  5. Faculty of Biology, Medicine, and Health, University of Manchester, Manchester, UK

    William Stuart

  6. The Experimental High School Affiliated to Beijing Normal University, Beijing, China

    Xiaochu Zhao

  7. The High School Affiliated to Renmin University, Beijing, China

    Zirui Yang

  8. National Laboratory of Macromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing, China

    Yuna Sun & Zihe Rao

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  1. Ran Zhang
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Contributions

Z.L., W.D. and Z.R. conceived the project. Z.L. and W.D. designed the experiments. Ra.Z., L.C., M.C., Z.S., M.H., Ro.Z., W.S., X.Z. and Z.Y. performed virus and protein purification, cryo-EM data collection and processing. Ra.Z., L.C., X.L., Y.S., S.L., W.D. and Z.L. analysed the data. Z.L., W.D. and Z.R. wrote the manuscript. All authors discussed the experiments, read and approved the manuscript.

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Correspondence to Wei Ding or Zhiyong Lou.

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Zhang, R., Cao, L., Cui, M. et al. Adeno-associated virus 2 bound to its cellular receptor AAVR. Nat Microbiol 4, 675–682 (2019). https://doi.org/10.1038/s41564-018-0356-7

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