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Nucleic Acids Research Advance Access published online on December 10, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm1080
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© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

RNA

Epstein–Barr virus-encoded microRNA miR-BART2 down-regulates the viral DNA polymerase BALF5

Stephanie Barth1, Thorsten Pfuhl1, Alfredo Mamiani1, Claudia Ehses1, Klaus Roemer2, Elisabeth Kremmer3, Christoph Jäker4, Julia Höck4, Gunter Meister4 and Friedrich A. Grässer1,*

1Institute of Virology, 2Jose-Carreras-Center Oncology Lab, University of Saarland Medical School, 66424 Homburg, 3Institute for Molecular Immunology, GSF, National Research Institute for Environment and Health, 81377 München and 4Laboratory of RNA Biology, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany

*To whom correspondence should be addressed. Tel: +49 6841 162 3983; Fax: +49 6841 162 3980; Email: graesser{at}uniklinik-saarland.de

Received August 20, 2007. Revised November 15, 2007. Accepted November 16, 2007.

MicroRNAs (miRNAs) have been implicated in sequence-specific cleavage, translational repression or deadenylation of specific target mRNAs resulting in post-transcriptional gene silencing. Epstein–Barr virus (EBV) encodes 23 miRNAs of unknown function. Here we show that the EBV-encoded miRNA miR-BART2 down-regulates the viral DNA polymerase BALF5. MiR-BART2 guides cleavage within the 3'-untranslated region (3'UTR) of BALF5 by virtue of its complete complementarity to its target. Induction of the lytic viral replication cycle results in a reduction of the level of miR-BART2 with a strong concomitant decrease of cleavage of the BALF5 3'UTR. Expression of miR-BART2 down-regulates the activity of a luciferase reporter gene containing the BALF5 3'UTR. Forced expression of miR-BART2 during lytic replication resulted in a 40–50% reduction of the level of BALF5 protein and a 20% reduction of the amount of virus released from EBV-infected cells. Our results are compatible with the notion that EBV-miR-BART2 inhibits transition from latent to lytic viral replication.

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.


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