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| The EMBO Journal
(2001) 20, 3282–3291, doi:10.1093/emboj/20.12.3282 |
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| DNA helicase-mediated packaging of adeno-associated virus type 2 genomes into preformed capsids |
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| Jason A. King2, Ralf Dubielzig3, Dirk Grimm1 and Jürgen A. Kleinschmidt1 |
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1 Applied Tumour Virology Program, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany 2 Present address: MRC Centre for Inflammation Research, Edinburgh University, Edinburgh, UK 3 Present address: Medigene AG, Lochhamer Stra  e 11, D-82152 Martinsried, GermanyTo whom correspondence should be addressed Jürgen A. Kleinschmidt, j.kleinschmidt@dkfz.de Received 5 March 2001; Revised 25 April 2001; Accepted 27 April 2001. |
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| Abstract |
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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. |
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| Keywords: AAV, DNA helicase, encapsidation, virus |
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