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PMID: 8016070 Published · ppublish English Journal Article

Adeno-associated virus (AAV) Rep proteins mediate complex formation between AAV DNA and its integration site in human DNA.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 91 ·No. 13 ·1994-06-21 ·Pages 5808-12

Weitzman MD, Kyöstiö SR, Kotin RM, Owens RA

Abstract

AAV is unique among eukaryotic viruses in the ability of its DNA to integrate preferentially into a specific region of the human genome. Understanding AAV integration may aid in developing gene therapy systems with predictable integration sites. Using a gel mobility-shift assay, we have identified a DNA sequence within the AAV integration locus on human chromosome 19 which is specifically bound by the AAV Rep78 and Rep68 proteins. This Rep recognition sequence is a GCTC repeating motif very similar to sequences within the inverted terminal repeats of the AAV genome which are also bound by Rep78 and Rep68. Cloned oligonucleotides containing the recognition sequence can direct specific binding by Rep proteins. Binding assays with mutant Rep proteins show that the amino-terminal portion of Rep78 and Rep68 can direct binding to either the AAV terminal repeat hairpin DNA or chromosome 19. This human genomic DNA can be complexed with AAV DNA by Rep proteins as demonstrated by a dual-label (32P/biotin) assay. These results suggest a role for Rep in targeting viral integration.

MeSH Terms
Bacterial Proteins/isolation & purification,metabolism Base Sequence Binding Sites Binding, Competitive Chromosomes, Human, Pair 19 Cloning, Molecular DNA/chemistry,genetics,metabolism DNA Helicases DNA, Viral/chemistry,genetics,metabolism DNA-Binding Proteins/isolation & purification,metabolism Dependovirus/genetics,metabolism Genome, Viral Humans Methylation Models, Structural Molecular Sequence Data Nucleic Acid Conformation Oligodeoxyribonucleotides Recombinant Proteins/isolation & purification,metabolism Trans-Activators Virus Integration
Chemicals
Bacterial Proteins DNA, Viral DNA-Binding Proteins Oligodeoxyribonucleotides Recombinant Proteins Trans-Activators replication initiator protein DNA DNA Helicases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Weitzman M D
Laboratory of Molecular and Cellular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892.
Kyöstiö S R
Kotin R M
Owens R A
References (26)
26 references, click to expand
  1. Organization of adeno-associated virus DNA in latently infected Detroit 6 cells.
    Virology. 1989 Jun;170(2):460-7 PMID: 2543124
  2. Factors that bind to adeno-associated virus terminal repeats.
    J Virol. 1989 Jul;63(7):3095-104 PMID: 2542617
  3. Site-specific integration by adeno-associated virus.
    Proc Natl Acad Sci U S A. 1990 Mar;87(6):2211-5 PMID: 2156265
  4. The AAV origin binding protein Rep68 is an ATP-dependent site-specific endonuclease with DNA helicase activity.
    Cell. 1990 May 4;61(3):447-57 PMID: 2159383
  5. Repair of deletions and double-strand gaps by homologous recombination in a mammalian in vitro system.
    Mol Cell Biol. 1991 Jan;11(1):445-57 PMID: 1986239
  6. Adeno-associated virus rep proteins produced in insect and mammalian expression systems: wild-type and dominant-negative mutant proteins bind to the viral replication origin.
    Virology. 1991 Sep;184(1):14-22 PMID: 1651588
  7. Mapping and direct visualization of a region-specific viral DNA integration site on chromosome 19q13-qter.
    Genomics. 1991 Jul;10(3):831-4 PMID: 1653762
  8. Targeted integration of adeno-associated virus (AAV) into human chromosome 19.
    EMBO J. 1991 Dec;10(12):3941-50 PMID: 1657596
  9. Gene transfer in human lymphocytes using a vector based on adeno-associated virus.
    J Immunother (1991). 1992 May;11(4):231-7 PMID: 1318074
  10. Regulated high level expression of a human gamma-globin gene introduced into erythroid cells by an adeno-associated virus vector.
    Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7257-61 PMID: 1323131
  11. Gene expression from adeno-associated virus vectors in airway epithelial cells.
    Am J Respir Cell Mol Biol. 1992 Sep;7(3):349-56 PMID: 1325813
  12. Dual-target inhibition of HIV-1 in vitro by means of an adeno-associated virus antisense vector.
    Science. 1992 Nov 27;258(5087):1485-8 PMID: 1359646
  13. Characterization of a preferred site on human chromosome 19q for integration of adeno-associated virus DNA by non-homologous recombination.
    EMBO J. 1992 Dec;11(13):5071-8 PMID: 1334463
  14. Identification of a DNA-binding domain in the amino terminus of adeno-associated virus Rep proteins.
    J Virol. 1993 Feb;67(2):997-1005 PMID: 8380475
  15. Expression of the cystic fibrosis transmembrane conductance regulator from a novel adeno-associated virus promoter.
    J Biol Chem. 1993 Feb 15;268(5):3781-90 PMID: 7679117
  16. Features of the adeno-associated virus origin involved in substrate recognition by the viral Rep protein.
    J Virol. 1993 Oct;67(10):6096-104 PMID: 8396670
  17. Biologically active Rep proteins of adeno-associated virus type 2 produced as fusion proteins in Escherichia coli.
    J Virol. 1994 Feb;68(2):797-804 PMID: 8289383
  18. Detection of adeno-associated virus (AAV)-specific nucleotide sequences in DNA isolated from latently infected Detroit 6 cells.
    Virology. 1975 Dec;68(2):556-60 PMID: 1198930
  19. Origin and termination of adeno-associated virus DNA replication.
    Virology. 1977 May 15;78(2):488-99 PMID: 867815
  20. Nucleotide sequence and organization of the adeno-associated virus 2 genome.
    J Virol. 1983 Feb;45(2):555-64 PMID: 6300419
  21. Genetics of adeno-associated virus: isolation and preliminary characterization of adeno-associated virus type 2 mutants.
    J Virol. 1984 Aug;51(2):329-39 PMID: 6086948
  22. Latent infection of KB cells with adeno-associated virus type 2.
    J Virol. 1986 Nov;60(2):515-24 PMID: 3021985
  23. Identification of the trans-acting Rep proteins of adeno-associated virus by antibodies to a synthetic oligopeptide.
    J Virol. 1986 Dec;60(3):823-32 PMID: 3023672
  24. Sequence requirements of Escherichia coli attTn7, a specific site of transposon Tn7 insertion.
    J Bacteriol. 1988 Jan;170(1):352-8 PMID: 2826397
  25. Identification of nuclear proteins that specifically interact with adeno-associated virus type 2 inverted terminal repeat hairpin DNA.
    J Virol. 1989 Jul;63(7):3034-9 PMID: 2542611
  26. Replication of a human parvovirus nonsense mutant in mammalian cells containing an inducible amber suppressor.
    Virology. 1989 Jul;171(1):239-47 PMID: 2545030
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.91.13.5808
Published
1994-06-21
Pages
5808-12
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC44086
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.91.13.5808 PMC Full Text View journal details
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