Document Detail


Alpha2,3 and alpha2,6 N-linked sialic acids facilitate efficient binding and transduction by adeno-associated virus types 1 and 6.
MedLine Citation:
PMID:  16940521     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Recombinant adeno-associated viruses (AAVs) are promising vectors in the field of gene therapy. Different AAV serotypes display distinct tissue tropism, believed to be related to the distribution of their receptors on target cells. Of the 11 well-characterized AAV serotypes, heparan sulfate proteoglycan and sialic acid have been suggested to be the attachment receptors for AAV type 2 and types 4 and 5, respectively. In this report, we identify the receptor for the two closely related serotypes, AAV1 and AAV6. First, we demonstrate using coinfection experiments and luciferase reporter analysis that AAV1 and AAV6 compete for similar receptors. Unlike heparin sulfate, enzymatic or genetic removal of sialic acid markedly reduced AAV1 and AAV6 binding and transduction. Further analysis using lectin staining and lectin competition assays identified that AAV1 and AAV6 use either alpha2,3-linked or alpha2,6-linked sialic acid when transducing numerous cell types (HepG2, Pro-5, and Cos-7). Treatment of cells with proteinase K but not glycolipid inhibitor reduced AAV1 and AAV6 infection, supporting the hypothesis that the sialic acid that facilitates infection is associated with glycoproteins rather than glycolipids. In addition, we determined by inhibitor (N-benzyl GalNAc)- and cell line-specific (Lec-1) studies that AAV1 and AAV6 require N-linked and not O-linked sialic acid. Furthermore, a resialylation experiment on a deficient Lec-2 cell line confirmed a 2,3 and 2,6 N-linked sialic acid requirement, while studies of mucin with O-linked sialic acid showed no inhibition effect for AAV1 and AAV6 transduction on Cos-7 cells. Finally, using a glycan array binding assay we determined that AAV1 efficiently binds to NeuAcalpha2-3GalNAcbeta1-4GlcNAc, as well as two glycoproteins with alpha2,3 and alpha2,6 N-linked sialic acids. Taken together, competition, genetic, inhibitor, enzymatic reconstitution, and glycan array experiments support alpha2,3 and alpha2,6 sialic acids that are present on N-linked glycoproteins as primary receptors for efficient AAV1 and AAV6 viral infection.
Authors:
Zhijian Wu; Edward Miller; Mavis Agbandje-McKenna; Richard Jude Samulski
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of virology     Volume:  80     ISSN:  0022-538X     ISO Abbreviation:  J. Virol.     Publication Date:  2006 Sep 
Date Detail:
Created Date:  2006-08-30     Completed Date:  2006-10-20     Revised Date:  2013-06-07    
Medline Journal Info:
Nlm Unique ID:  0113724     Medline TA:  J Virol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  9093-103     Citation Subset:  IM    
Affiliation:
Gene Therapy Center, CB # 7352, 7119 Thurston Building, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7352, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
COS Cells
Cell Line
Cell Membrane / metabolism
Cercopithecus aethiops
Dependovirus / metabolism*
Endopeptidase K / chemistry
Glycolipids / chemistry
Heparan Sulfate Proteoglycans / metabolism
Humans
Protein Binding
Recombinant Proteins / chemistry
Sialic Acids / chemistry,  metabolism,  physiology*
Grant Support
ID/Acronym/Agency:
GM 059299/GM/NIGMS NIH HHS; GM 62116/GM/NIGMS NIH HHS; HL 051818/HL/NHLBI NIH HHS; HL 069973/HL/NHLBI NIH HHS; HL 51811/HL/NHLBI NIH HHS; HL 59412/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Glycolipids; 0/Heparan Sulfate Proteoglycans; 0/Recombinant Proteins; 0/Sialic Acids; EC 3.4.21.64/Endopeptidase K
Comments/Corrections

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