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Journal of Virology, December 2002, p. 12792-12802, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12792-12802.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

A cis-Acting Element That Directs Circular Adeno-Associated Virus Replication and Packaging

Sergei Musatov,1 Jill Roberts,2 Donald Pfaff,1 and Michael Kaplitt1,2*

Laboratory of Neurobiology and Behavior, The Rockefeller University,1 Department of Neurosurgery, Weill Medical College of Cornell University, New York, New York 100212

Received 18 March 2002/ Accepted 4 September 2002

A novel pathway of adeno-associated virus (AAV) replication marked by the assembly of circular monomer duplex intermediates (cAAV) has been recently discovered. In the present report we identify a single AD domain of the inverted terminal repeat as a minimal origin of cAAV replication. A small internal palindrome (BB'), necessary for optimal Rep-inverted terminal repeat interaction, does not contribute to the efficiency of cAAV replication, while the terminal resolution site is an essential cis-acting element. Furthermore, recombinant cAAV vectors that encompass only the AD domain replicate exclusively in a circular form and no detectable linear duplex replicative intermediates are generated, suggesting that both pathways of AAV replication are independent and can be separated. In addition, we show that cAAVs are efficient templates for encapsidation of single-stranded DNA genomes, an observation that assigns a biological role for these novel replication species. Together, these findings shed new light on the current model of AAV replication and packaging.

* Corresponding author. Mailing address: Weill Medical College of Cornell University, Department of Neurosurgery, 525 East 68 St., New York, NY 10021. Phone: (212) 746-5265. Fax: (212) 746-8226. E-mail: mik2002{at}med.cornell.edu.

Journal of Virology, December 2002, p. 12792-12802, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12792-12802.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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