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Journal of Virology, July 2006, p. 6855-6864, Vol. 80, No. 14
0022-538X/06/$08.00+0     doi:10.1128/JVI.00383-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Regulation of Adeno-Associated Virus DNA Replication by the Cellular TAF-I/Set Complex{dagger}

Gianluca Pegoraro,1 Alessandro Marcello,2 Michael P. Myers,3 and Mauro Giacca1*

Molecular Medicine Laboratory,1 Molecular Virology Laboratory, International Center for Genetic Engineering and Biotechnology, Trieste, Italy,2 Cold Spring Harbor Laboratory, Cold Spring Harbor, New York3

Received 23 February 2006/ Accepted 4 May 2006

The Rep proteins of the adeno-associated virus (AAV) are required for viral replication in the presence of adenovirus helper functions and as yet poorly characterized cellular factors. In an attempt to identify such factors, we purified Flag-Rep68-interacting proteins from human cell lysates. Several polypeptides were identified by mass spectrometry, among which was ANP32B, a member of the acidic nuclear protein 32 family which takes part in the formation of the template-activating factor I/Set oncoprotein (TAF-I/Set) complex. The N terminus of Rep was found to specifically bind the acidic domain of ANP32B; through this interaction, Rep was also able to recruit other members of the TAF-I/Set complex, including the ANP32A protein and the histone chaperone TAF-I/Set. Further experiments revealed that silencing of ANP32A and ANP32B inhibited AAV replication, while overexpression of all of the components of the TAF-I/Set complex increased de novo AAV DNA synthesis in permissive cells. Besides being the first indication that the TAF-I/Set complex participates in wild-type AAV replication, these findings have important implications for the generation of recombinant AAV vectors since overexpression of the TAF-I/Set components was found to markedly increase viral vector production.

* Corresponding author. Mailing address: ICGEB Trieste, Padriciano, 99, 34012 Trieste, Italy. Phone: 39-040-3757-324. Fax: 39-040-226-555. E-mail: giacca{at}icgeb.org.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, July 2006, p. 6855-6864, Vol. 80, No. 14
0022-538X/06/$08.00+0     doi:10.1128/JVI.00383-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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