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Journal of Virology, July 2006, p. 6873-6882, Vol. 80, No. 14
0022-538X/06/$08.00+0     doi:10.1128/JVI.00275-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Experimental Confirmation of Global Murine Cytomegalovirus Open Reading Frames by Transcriptional Detection and Partial Characterization of Newly Described Gene Products{dagger}

Qiyi Tang,1 Eain A. Murphy,2 and Gerd G. Maul1*

The Wistar Institute, 3601 Spruce Street, Philadelphia, Pennsylvania 19104,1 Department of Molecular Biology, Princeton University, Princeton, New Jersey 805442

Received 7 February 2006/ Accepted 1 May 2006

Murine cytomegalovirus (MCMV) and human CMV (HCMV) share many features making the mouse system a potential small-animal model for HCMV. Although the genomic DNA sequence and the predicted open reading frames (ORFs) of MCMV have been determined, experimental evidence that the ORFs are actually transcribed has been lacking. We developed an MCMV global-DNA microarray that includes all previously predicted ORFs and 14 potential ones. A total of 172 ORFs were confirmed to be transcribed, including 7 newly discovered ORFs not previously predicted. No gene products from 10 previously predicted ORFs were detected by either DNA microarray analysis or reverse transcriptase PCR in MCMV-infected mouse fibroblasts, although 2 of those were expressed in a macrophage cell line, suggesting that potential gene products from these open reading fames are silenced in fibroblasts and required in macrophages. Immunohistochemical localization of the six newly described ORF products and three recently identified ones in cells transfected with the respective construct revealed four of the products in the nucleus and five in mitochondria. Analysis of two ORFs using site-directed mutagenesis showed that deletion of one of the mitochondrion-localized gene products led to significantly decreased replication in fibroblasts.

* Corresponding author. Mailing address: The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104. Phone: (215) 898-3817. E-mail: maul{at}wistar.org.

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

Journal of Virology, July 2006, p. 6873-6882, Vol. 80, No. 14
0022-538X/06/$08.00+0     doi:10.1128/JVI.00275-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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