Characterization of a preferred site on human chromosome 19q for integration of adeno-associated virus DNA by non-homologous recombination.
The human parvovirus, adeno-associated virus (AAV), has been shown to integrate preferentially into human chromosome 19 q13.3-qter. The human target sequence for AAV integration (AAVS1) was cloned and sequenced. By analysis of the proviral junctions it was determined that integration of the AAV DNA occurred via a non-homologous recombination pathway although there were either four or five identical nucleotides at the junctions. Integration was a multistep, concerted process that resulted in cellular sequence rearrangements. The sequence of the integration locus was analyzed for possible recombination signals. Direct repeats at a much greater than random occurrence were found distributed non-uniformly throughout the AAVS1 sequence. A CpG island containing transcription factor binding site elements is suggestive of a TATA-less promoter. Evidence for transcriptional activity was provided by PCR amplification of reverse transcribed RNA.
Kotin
R M
RM
Molecular Hematology Branch, National Institutes of Health, Bethesda, MD 20892.
Linden
R M
RM
Berns
K I
KI
D001483:Base Sequence; D002888:Chromosomes, Human, Pair 19; D003001:Cloning, Molecular; D004247:DNA; D004279:DNA, Viral / Q000378:metabolism*; D000229:Dependovirus / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D016385:TATA Box; D014158:Transcription, Genetic; D016662:Virus Integration
D004279:DNA, Viral; D004247:DNA