Adenoassociated virus (AAV) serotypes have distinctive interactions with domains of the cellular AAV receptor

Sirika Pillay, Wei Zou, Fang Cheng, Andreas S. Puschnik, Nancy L. Meyer, Safder S. Ganaie, Xuefeng Deng, Jonathan E. Wosen, Omar Davulcu, Ziying Yan, John F. Engelhardt, Kevin E. Brown, Michael Chapman, Jianming Qiu, Jan E. Carette

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Adeno-associated virus (AAV) entry is determined by its interactions with specific surface glycans and a proteinaceous receptor(s). Adeno-associated virus receptor (AAVR) (also named KIAA0319L) is an essential cellular receptor required for the transduction of vectors derived from multiple AAV serotypes, including the evolutionarily distant serotypes AAV2 and AAV5. Here, we further biochemically characterize the AAV-AAVR interaction and define the domains within the ectodomain of AAVR that facilitate this interaction. By using a virus overlay assay, it was previously shown that the major AAV2 binding protein in membrane preparations of human cells corresponds to a glycoprotein with a molecular mass of 150 kDa. By establishing a purification procedure, performing further protein separation by two-dimensional electrophoresis, and utilizing mass spectrometry, we now show that this glycoprotein is identical to AAVR. While we find that AAVR is an N-linked glycosylated protein, this glycosylation is not a strict requirement for AAV2 binding or functional transduction. Using a combination of genetic complementation with deletion constructs and virus overlay assays with individual domains, we find that AAV2 functionally interacts predominantly with the second Ig-like polycystic kidney disease (PKD) repeat domain (PKD2) present in the ectodomain of AAVR. In contrast, AAV5 interacts primarily through the first, most membrane-distal, PKD domain (PKD1) of AAVR to promote transduction. Furthermore, other AAV serotypes, including AAV1 and -8, require a combination of PKD1 and PKD2 for optimal transduction. These results suggest that despite their shared dependence on AAVR as a critical entry receptor, different AAV serotypes have evolved distinctive interactions with the same receptor.

Original languageEnglish (US)
Article numbere00391-17
JournalJournal of Virology
Volume91
Issue number18
DOIs
StatePublished - Sep 1 2017

Fingerprint

Dependovirus
Virus Receptors
serotypes
Viruses
viruses
Polycystic Kidney Diseases
receptors
glycoproteins
Glycoproteins
genetic complementation
virus receptors
Serogroup
Virus Internalization
Membranes
assays
glycosylation
Glycosylation
electrophoresis
Polysaccharides
Electrophoresis

Keywords

  • AAVR
  • Adeno-associated virus
  • Gene therapy
  • Receptor-ligand interaction
  • Viral receptor
  • Virus overlay assay
  • Virus-host interactions

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Pillay, S., Zou, W., Cheng, F., Puschnik, A. S., Meyer, N. L., Ganaie, S. S., ... Carette, J. E. (2017). Adenoassociated virus (AAV) serotypes have distinctive interactions with domains of the cellular AAV receptor. Journal of Virology, 91(18), [e00391-17]. https://doi.org/10.1128/JVI.00391-17

Adenoassociated virus (AAV) serotypes have distinctive interactions with domains of the cellular AAV receptor. / Pillay, Sirika; Zou, Wei; Cheng, Fang; Puschnik, Andreas S.; Meyer, Nancy L.; Ganaie, Safder S.; Deng, Xuefeng; Wosen, Jonathan E.; Davulcu, Omar; Yan, Ziying; Engelhardt, John F.; Brown, Kevin E.; Chapman, Michael; Qiu, Jianming; Carette, Jan E.

In: Journal of Virology, Vol. 91, No. 18, e00391-17, 01.09.2017.

Research output: Contribution to journalArticle

Pillay, S, Zou, W, Cheng, F, Puschnik, AS, Meyer, NL, Ganaie, SS, Deng, X, Wosen, JE, Davulcu, O, Yan, Z, Engelhardt, JF, Brown, KE, Chapman, M, Qiu, J & Carette, JE 2017, 'Adenoassociated virus (AAV) serotypes have distinctive interactions with domains of the cellular AAV receptor', Journal of Virology, vol. 91, no. 18, e00391-17. https://doi.org/10.1128/JVI.00391-17
Pillay, Sirika ; Zou, Wei ; Cheng, Fang ; Puschnik, Andreas S. ; Meyer, Nancy L. ; Ganaie, Safder S. ; Deng, Xuefeng ; Wosen, Jonathan E. ; Davulcu, Omar ; Yan, Ziying ; Engelhardt, John F. ; Brown, Kevin E. ; Chapman, Michael ; Qiu, Jianming ; Carette, Jan E. / Adenoassociated virus (AAV) serotypes have distinctive interactions with domains of the cellular AAV receptor. In: Journal of Virology. 2017 ; Vol. 91, No. 18.
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