Amino-terminal domain exchange redirects origin-specific interactions of adeno-associated virus Rep78 in vitro

M. Yoon, D. H. Smith, P. Ward, F. J. Medrano, A. K. Aggarwal, R. M. Linden

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The unique ability of adeno-associated virus type 2 (AAV) to site-specifically integrate its genome into a defined sequence on human chromosome 19 (AAVS1) makes it of particular interest for use in targeted gene delivery. The objective underlying this study is to provide evidence for the feasibility of retargeting site-specific integration into selected loci within the human genome. Current models postulate that AAV DNA integration is initiated through the interactions of the products of a single viral open reading frame, REP, with sequences present in AAVS1 that resemble the minimal origin for AAV DNA replication. Here, we present a cell-free system designed to dissect the Rep functions required to target site-specific integration using functional chimeric Rep proteins derived from AAV Rep78 and Rep1 of the closely related goose parvovirus. We show that amino-terminal domain exchange efficiently redirects the specificity of Rep to the minimal origin of DNA replication. Furthermore, we establish that the amino-terminal 208 amino acids of Rep78/68 constitute a catalytic domain of Rep sufficient to mediate site-specific endonuclease activity.

Original languageEnglish (US)
Pages (from-to)3230-3239
Number of pages10
JournalJournal of Virology
Volume75
Issue number7
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

Dependovirus
DNA Replication
DNA replication
Chromosomes, Human, Pair 19
Geese
Parvovirus
Replication Origin
Cell-Free System
Endonucleases
Human Chromosomes
Human Genome
recombinant fusion proteins
cell free system
Open Reading Frames
genome
Catalytic Domain
gene transfer
active sites
Genome
open reading frames

ASJC Scopus subject areas

  • Immunology

Cite this

Amino-terminal domain exchange redirects origin-specific interactions of adeno-associated virus Rep78 in vitro. / Yoon, M.; Smith, D. H.; Ward, P.; Medrano, F. J.; Aggarwal, A. K.; Linden, R. M.

In: Journal of Virology, Vol. 75, No. 7, 2001, p. 3230-3239.

Research output: Contribution to journalArticle

Yoon, M. ; Smith, D. H. ; Ward, P. ; Medrano, F. J. ; Aggarwal, A. K. ; Linden, R. M. / Amino-terminal domain exchange redirects origin-specific interactions of adeno-associated virus Rep78 in vitro. In: Journal of Virology. 2001 ; Vol. 75, No. 7. pp. 3230-3239.
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