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Journal of Virology, July 2006, p. 6973-6981, Vol. 80, No. 14
0022-538X/06/$08.00+0     doi:10.1128/JVI.00123-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The L4 22-Kilodalton Protein Plays a Role in Packaging of the Adenovirus Genome

Philomena Ostapchuk, Mary E. Anderson, Sharanya Chandrasekhar, and Patrick Hearing*

Department of Molecular Genetics and Microbiology, School of Medicine, Stony Brook University, Stony Brook, New York 11794

Received 18 January 2006/ Accepted 15 April 2006

Packaging of the adenovirus (Ad) genome into a capsid is absolutely dependent upon the presence of a cis-acting region located at the left end of the genome referred to as the packaging domain. The functionally significant sequences within this domain consist of at least seven similar repeats, referred to as the A repeats, which have the consensus sequence 5' TTTG-N8-CG 3'. In vitro and in vivo binding studies have demonstrated that the adenovirus protein IVa2 binds to the CG motif of the packaging sequences. In conjunction with IVa2, another virus-specific protein binds to the TTTG motifs in vitro. The efficient formation of these protein-DNA complexes in vitro was precisely correlated with efficient packaging activity in vivo. We demonstrate that the binding activity to the TTTG packaging sequence motif is the product of the L4 22-kDa open reading frame. Previously, no function had been ascribed to this protein. Truncation of the L4 22-kDa protein in the context of the viral genome did not reduce viral gene expression or viral DNA replication but eliminated the production of infectious virus. We suggest that the L4 22-kDa protein, in conjunction with IVa2, plays a critical role in the recognition of the packaging domain of the Ad genome that leads to viral DNA encapsidation. The L4 22-kDa protein is also involved in recognition of transcription elements of the Ad major late promoter.

* Corresponding author. Mailing address: Department of Molecular Genetics and Microbiology, School of Medicine, Stony Brook University, Stony Brook, NY 11794. Phone: (631) 632-8813. Fax: (631) 632-8891. E-mail: phearing{at}ms.cc.sunysb.edu.

Journal of Virology, July 2006, p. 6973-6981, Vol. 80, No. 14
0022-538X/06/$08.00+0     doi:10.1128/JVI.00123-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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