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  • The EMBO Journal (2001) 20, 3282 - 3291
  • doi:10.1093/emboj/20.12.3282

DNA helicase-mediated packaging of adeno-associated virus type 2 genomes into preformed capsids

Jason A. King2, Ralf Dubielzig3, Dirk Grimm1 and Jürgen A. Kleinschmidt1

  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 Stras zlige 11, D-82152 Martinsried, Germany

Correspondence to:

Jürgen A. Kleinschmidt, E-mail: j.kleinschmidt@dkfz.de

Received 5 March 2001; Accepted 27 April 2001; Revised 25 April 2001

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'right arrow5' 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.

  • Keywords:

    • AAV,
    • DNA helicase,
    • encapsidation,
    • virus