Interaction of penton base Arg-Gly-Asp motifs with integrins is crucial for adenovirus serotype 35 vector transduction in human hematopoietic cells.
Most subgroup B adenoviruses (Ads), including adenovirus (Ad) serotype 35 (Ad35), bind to human CD46 as a receptor; however, the infection processes of subgroup B Ads following attachment to CD46 remain to be elucidated. Subgroup B Ads possess Arg-Gly-Asp (RGD) motifs in the penton base, similarly to subgroup C Ad serotypes 2 and 5. In this study, we examined the role of penton base RGD motifs in Ad35 vector-mediated transduction in human hematopoietic cells. Inhibition of interaction between integrins and the RGD motifs by divalent cation chelation and a synthetic RGD peptide reduced the transduction efficiencies of Ad35 vectors; however, the amounts of cell-associated vector DNA of Ad35 vectors at 4 or 37 degrees C were not decreased by divalent cation chelation or the RGD peptide. Mutation of penton base RGD motifs reduced the transduction efficiencies of Ad35 vectors, although the amounts of cell-associated vector DNA of Ad35 vectors at 4 or 37 degrees C were not altered by mutation of penton base RGD motifs in Ad35 vectors. Furthermore, preincubation with several types of anti-integrin antibodies significantly inhibited Ad35 vector-mediated transduction. These results suggest that interaction between integrins and penton base RGD motifs plays a crucial role in Ad35 vector-mediated transduction in hematopoietic cells, probably in the post-internalization steps.
Murakami
S
S
Laboratory of Gene Transfer and Regulation, National Institute of Biomedical Innovation, Ibaraki City, Osaka, Japan.
Sakurai
F
F
Kawabata
K
K
Okada
N
N
Fujita
T
T
Yamamoto
A
A
Hayakawa
T
T
Mizuguchi
H
H
D000256:Adenoviridae / Q000235:genetics*; D020816:Amino Acid Motifs; D036022:Capsid Proteins / Q000235:genetics*; D002460:Cell Line; D002478:Cells, Cultured; D005434:Flow Cytometry; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000821:virology*; D006801:Humans; D016023:Integrins / Q000032:analysis / Q000235:genetics*; D009154:Mutation; D009842:Oligopeptides / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods; D016662:Virus Integration