Identification and Mutagenesis of the Adeno-Associated Virus 5 Sialic Acid Binding Region

作者：Sandra Afione, Michael A. DiMattia, Sujata Halder, Giovanni Di Pasquale, Mavis Agbandje‐McKenna, John A. Chiorini · 发表于：Journal of Virology · 年份：2014 · DOI：10.1128/jvi.02503-14 · 被引用次数：56 · 研究领域：Virus-based gene therapy research、Viral gastroenteritis research and epidemiology、Cytomegalovirus and herpesvirus research

UNLABELLED: 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 ...