Format

Send to

Choose Destination
J Virol. 2015 Feb;89(3):1660-72. doi: 10.1128/JVI.02503-14. Epub 2014 Nov 19.

Identification and mutagenesis of the adeno-associated virus 5 sialic acid binding region.

Author information

1
NIDCR, NIH, Molecular Physiology and Therapeutics Branch, Bethesda, Maryland, USA.
2
University of Florida, Department of Biochemistry and Molecular Biology, Gainesville, Florida, USA.
3
University of Florida, Department of Biochemistry and Molecular Biology, Gainesville, Florida, USA mckenna@ufl.edu jchiorini@dir.nidcr.nih.gov.
4
NIDCR, NIH, Molecular Physiology and Therapeutics Branch, Bethesda, Maryland, USA mckenna@ufl.edu jchiorini@dir.nidcr.nih.gov.

Abstract

As a genus, the dependoviruses use a diverse group of cell surface carbohydrates for attachment and entry. Despite the fact that a majority of adeno-associated viruses (AAVs) utilize sialic acid (SIA) for binding and transduction, this virus-carbohydrate interaction is poorly understood. Utilizing X-ray crystallography, two SIA binding regions were mapped for AAV5. The first site mapped to the depression in the center of the 3-fold axis of symmetry, while the second site was located under the βHI loop close to the 5-fold axis. Mutagenesis of amino acids 569 and 585 or 587 within the 3-fold depression resulted in elimination or alteration in SIA-dependent transduction, respectively. This change in SIA binding was confirmed using glycan microarrays. Mutagenesis of the second site identified a role in transduction that was SIA independent. Further studies of the mutants at the 3-fold site demonstrated a change in transduction activity and cell tropism in vivo as well as resistance to neutralization by a polyclonal antibody raised against the wild-type virus.

IMPORTANCE:

Despite the fact that a majority of AAVs utilize sialic acid for binding and transduction, this virus-carbohydrate interaction is poorly understood. Utilizing X-ray crystallography, the sialic acid binding regions of AAV5 were identified and studied using a variety of approaches. Mutagenesis of this region resulted in elimination or alteration in sialic acid-dependent transduction in cell lines. This change in sialic acid glycan binding was confirmed using glycan arrays. Further study also demonstrated a change in transduction and activity and cell tropism in vivo as well as resistance to neutralization by antibodies raised against the wild-type virus.

PMID:
25410855
PMCID:
PMC4300766
DOI:
10.1128/JVI.02503-14
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center