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Nucleic Acids Research 2006 34(1):353-363; doi:10.1093/nar/gkj440
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Published online 12 January 2006

© The Author 2006. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oxfordjournals.org

Article

The hepatitis B virus PRE contains a splicing regulatory element

Tilman Heise*, Gunhild Sommer, Kerstin Reumann, Imke Meyer1, Hans Will and Heiner Schaal1

Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie an der Universität Hamburg Martinistrasse 52, D-20251 Hamburg, Germany 1Institut für Virologie, Universitätsstrasse 1 40225 Düsseldorf, Germany

*To whom correspondence should be addressed. Tel: +49 40 48051 220; Fax: +49 40 48051 222; Email: heise{at}hpi.uni-hamburg.de

Received October 20, 2005. Revised December 12, 2005. Accepted December 23, 2005.

The posttranscriptional regulatory element (PRE) is considered to enhance hepatitis B virus (HBV) gene expression by facilitating the nuclear export of intronless viral subgenomic RNAs. Its role in the RNA metabolism of the viral pregenomic RNA (pgRNA) is currently unknown. We identified a positively cis-acting splicing regulatory element (SRE-1) and present two lines of evidence for its functionality. Firstly, in a heterologous context SRE-1 functionally substitutes for a retroviral bidirectional exonic splicing enhancer (ESE). As expected, SRE-1 is a splicing enhancer also in its natural viral sequence context, since deletion of SRE-1 reduces splicing of pgRNA in cell culture experiments. Secondly, we show that stimulation of HBV RNA splicing by the splicing factor PSF was repressed by the PRE. Analysis of a variety of PSF mutants indicated that RNA-binding and protein–protein interaction were required to enhance splicing. In addition, we show that the PRE contributed to pgRNA stability, but has little influence on its nuclear export. Herein, we report for the first time that the PRE harbors splicing stimulating and inhibiting regulatory elements controlling processing of the viral pregenome. We discuss a model in which the regulation of pgRNA splicing depends on cellular factors interacting with the PRE.


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M. Schwalbe, O. Ohlenschlager, A. Marchanka, R. Ramachandran, S. Hafner, T. Heise, and M. Gorlach
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