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Journal of Virology, December 2007, p. 13761-13770, Vol. 81, No. 24
0022-538X/07/$08.00+0     doi:10.1128/JVI.01290-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Discrete Clusters of Virus-Encoded MicroRNAs Are Associated with Complementary Strands of the Genome and the 7.2-Kilobase Stable Intron in Murine Cytomegalovirus{triangledown}

Amy H. Buck,1* Javier Santoyo-Lopez,1 Kevin A. Robertson,1 Diwakar S. Kumar,1 Martin Reczko,2 and Peter Ghazal1

Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom,1 Foundation for Research and Technology—Hellas (FORTH), Heraklion, Greece2

Received 13 June 2007/ Accepted 1 October 2007

The prevalence and importance of microRNAs (miRNAs) in viral infection are increasingly relevant. Eleven miRNAs were previously identified in human cytomegalovirus (HCMV); however, miRNA content in murine CMV (MCMV), which serves as an important in vivo model for CMV infection, has not previously been examined. We have cloned and characterized 17 novel miRNAs that originate from at least 12 precursor miRNAs in MCMV and are not homologous to HCMV miRNAs. In parallel, we applied a computational analysis, using a support vector machine approach, to identify potential precursor miRNAs in MCMV. Four of the top 10 predicted precursor sequences were cloned in this study, and the combination of computational and cloning analysis demonstrates that MCMV has the capacity to encode miRNAs clustered throughout the genome. On the basis of drug sensitivity experiments for resolving the kinetic class of expression, we show that the MCMV miRNAs are both early and late gene products. Notably, the MCMV miRNAs occur on complementary strands of the genome in specific regions, a feature which has not previously been observed for viral miRNAs. One cluster of miRNAs occurs in close proximity to the 5' splice site of the previously identified 7.2-kb stable intron, implying a variety of potential regulatory mechanisms for MCMV miRNAs.

* Corresponding author. Mailing address: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom. Phone: 44 131 242 6280. Fax: 44 131 242 6244. E-mail: a.buck{at}ed.ac.uk

{triangledown} Published ahead of print on 10 October 2007.

Journal of Virology, December 2007, p. 13761-13770, Vol. 81, No. 24
0022-538X/07/$08.00+0     doi:10.1128/JVI.01290-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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