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Journal of Virology, May 2007, p. 5349-5361, Vol. 81, No. 10
0022-538X/07/$08.00+0     doi:10.1128/JVI.02624-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Epigenetic Status of an Adenovirus Type 12 Transgenome upon Long-Term Cultivation in Hamster Cells{triangledown}

Norbert Hochstein,1 Indrikis Muiznieks,2,3 Laurence Mangel,2,{dagger} Holger Brondke,1,{ddagger} and Walter Doerfler1,2*

Institute for Clinical and Molecular Virology, Erlangen University Medical School, Schlossgarten 4, D-91054 Erlangen, Germany,1 Institute of Genetics, University of Cologne, Zülpicher Str. 47, D-50674 Cologne, Germany,2 Department of Microbiology, Latvijas Universitate, 19 Raina Blvd., LV-1586 Riga, Latvia3

Received 28 November 2006/ Accepted 27 February 2007

The epigenetic status of integrated adenovirus type 12 (Ad12) DNA in hamster cells cultivated for about 4 decades has been investigated. Cell line TR12, a fibroblastic revertant of the Ad12-transformed epitheloid hamster cell line T637 with 15 copies of integrated Ad12 DNA, carries one Ad12 DNA copy plus a 3.9-kbp fragment from a second copy. The cellular insertion site for the Ad12 integrate, identical in both cell lines, is a >5.2-kbp inverted DNA repeat. The Ad12 transgenome is packaged around nucleosomes. The cellular junction is more sensitive to micrococcal nuclease at Ad12-occupied sites than at unoccupied sites. Bisulfite sequencing reveals complete de novo methylation in most of the 1,634 CpGs of the integrated viral DNA, except for its termini. Isolated unmethylated CpGs extend over the entire Ad12 integrate. The fully methylated transgenome segments are characterized by promoter silencing and histone H3 and H4 hypoacetylation. Nevertheless, there is minimal transcriptional activity of the late viral genes controlled by the fully methylated major late promoter of Ad12 DNA.

* Corresponding author. Mailing address: Institute for Virology, Erlangen University Medical School, Schlossgarten 4, D-91054 Erlangen, Germany. Phone: (49) 9131-852-6002. Fax: (49) 9131-852-2101. E-mail: walter.doerfler{at}viro.med.uni-erlangen.de

{triangledown} Published ahead of print on 7 March 2007.

{dagger} Present address: amaxa GmbH, Nattermannallee 1, D-50829 Cologne, Germany.

{ddagger} Present address: Universitätsklinikum Bonn, Sigmund-Freud-Str. 25, D-53127 Bonn, Germany.

Journal of Virology, May 2007, p. 5349-5361, Vol. 81, No. 10
0022-538X/07/$08.00+0     doi:10.1128/JVI.02624-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Hochstein, N., Webb, D., Hosel, M., Seidel, W., Auerochs, S., Doerfler, W. (2008). Human CAR Gene Expression in Nonpermissive Hamster Cells Boosts Entry of Type 12 Adenovirions and Nuclear Import of Viral DNA. J. Virol. 82: 4159-4163 [Abstract] [Full Text]  


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