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Journal of Virology, December 2007, p. 12918-12926, Vol. 81, No. 23
0022-538X/07/$08.00+0     doi:10.1128/JVI.01531-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification of a New Human Adenovirus Protein Encoded by a Novel Late l-Strand Transcription Unit{triangledown}

Ann E. Tollefson,1 Baoling Ying,2 Konstantin Doronin,1,{dagger} Peter D. Sidor,3 and William S. M. Wold1*

Department of Molecular Microbiology and Immunology, Saint Louis University Health Sciences Center, 1402 South Grand Blvd., St. Louis, Missouri 63104,1 VirRx, Inc., 3681 Park Ave., St. Louis, Missouri 63110,2 School of Medicine, Saint Louis University, 1402 S. Grand Blvd., St. Louis, Missouri 631043

Received 12 July 2007/ Accepted 10 September 2007

A short open reading frame named the "U exon," located on the adenovirus (Ad) l-strand (for leftward transcription) between the early E3 region and the fiber gene, is conserved in mastadenoviruses. We have observed that Ad5 mutants with large deletions in E3 that infringe on the U exon display a mild growth defect, as well as an aberrant Ad E2 DNA-binding protein (DBP) intranuclear localization pattern and an apparent failure to organize replication centers during late infection. Mutants in which the U exon DNA is reconstructed have a reversed phenotype. Chow et al. (L. T. Chow et al., J. Mol. Biol. 134:265-303, 1979) described mRNAs initiating in the region of the U exon and spliced to downstream sequences in the late DBP mRNA leader and the DBP-coding region. We have cloned this mRNA (as cDNA) from Ad5 late mRNA; the predicted protein is 217 amino acids, initiating in the U exon and continuing in frame in the DBP leader and in the DBP-coding region but in a different reading frame from DBP. Polyclonal and monoclonal antibodies generated against the predicted U exon protein (UXP) showed that UXP is ~24K in size by immunoblot and is a late protein. At 18 to 24 h postinfection, UXP is strongly associated with nucleoli and is found throughout the nucleus; later, UXP is associated with the periphery of replication centers, suggesting a function relevant to Ad DNA replication or RNA transcription. UXP is expressed by all four species C Ads. When expressed in transient transfections, UXP complements the aberrant DBP localization pattern of UXP-negative Ad5 mutants. Our data indicate that UXP is a previously unrecognized protein derived from a novel late l-strand transcription unit.

* Corresponding author. Mailing address: Department of Molecular Microbiology and Immunology, St. Louis University Health Sciences Center, 1100 South Grand Blvd., St. Louis, MO 63104. Phone: (314) 977-8857. Fax: (314) 977-8717. E-mail: woldws{at}slu.edu

{triangledown} Published ahead of print on 19 September 2007.

{dagger} Present address: Division of Infectious Diseases, Mayo Clinic, Guggenheim 5-16, 200 First St. SW, Rochester, MN 55902.

Journal of Virology, December 2007, p. 12918-12926, Vol. 81, No. 23
0022-538X/07/$08.00+0     doi:10.1128/JVI.01531-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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