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DNA helicase‐mediated packaging of adeno‐associated virus type 2 genomes into preformed capsids
Author(s) -
King Jason A.,
Dubielzig Ralf,
Grimm Dirk,
Kleinschmidt Jürgen A.
Publication year - 2001
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/20.12.3282
Subject(s) - helicase , capsid , biology , processivity , dna , nucleic acid , genome , rna helicase a , circular bacterial chromosome , virology , adeno associated virus , microbiology and biotechnology , genetics , virus , dna replication , gene , rna , vector (molecular biology) , recombinant dna
Helicases not only catalyse the disruption of hydrogen bonding between complementary regions of nucleic acids, but also move along nucleic acid strands in a polar fashion. Here we show that the Rep52 and Rep40 proteins of adeno‐associated virus type 2 (AAV‐2) are required to translocate capsid‐associated, single‐stranded DNA genomes into preformed empty AAV‐2 capsids, and that the DNA helicase function of Rep52/40 is essential for this process. Furthermore, DNase protection experiments suggest that insertion of AAV‐2 genomes proceeds from the 3′ end, which correlates with the 3′→5′ processivity demonstrated for the Rep52/40 helicase. A model is proposed in which capsid‐immobilized helicase complexes act as molecular motors to ‘pump’ single‐stranded DNA across the capsid boundary.