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Journal of Virology, September 2002, p. 8532-8539, Vol. 76, No. 17
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.17.8532-8539.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Ebola Virus VP30-Mediated Transcription Is Regulated by RNA Secondary Structure Formation

Michael Weik, Jens Modrof, Hans-Dieter Klenk, Stephan Becker, and Elke Mühlberger*

Institut für Virologie der Philipps-Universität Marburg, 35037 Marburg, Germany

Received 25 March 2002/ Accepted 6 June 2002

The nucleocapsid protein VP30 of Ebola virus (EBOV), a member of the Filovirus family, is known to act as a transcription activator. By using a reconstituted minigenome system, the role of VP30 during transcription was investigated. We could show that VP30-mediated transcription activation is dependent on formation of a stem-loop structure at the first gene start site. Destruction of this secondary structure led to VP30-independent transcription. Analysis of the transcription products of bicistronic minigenomes with and without the ability to form the secondary structure at the first transcription start signal revealed that transcription initiation at the first gene start site is a prerequisite for transcription of the second gene, independent of the presence of VP30. When the transcription start signal of the second gene was exchanged with the transcription start signal of the first gene, transcription of the second gene also was regulated by VP30, indicating that the stem-loop structure of the first transcription start site acts autonomously and independently of its localization on the RNA genome. Our results suggest that VP30 regulates a very early step of EBOV transcription, most likely by inhibiting pausing of the transcription complex at the RNA structure of the first transcription start site.

* Corresponding author. Mailing address: Institut für Virologie der Philipps-Universität Marburg, Robert-Koch-Str. 17, 35037 Marburg, Germany. Phone: 496421-2864314. Fax: 496421-2865482. E-mail: muehlber{at}mailer.uni-marburg.de.

Journal of Virology, September 2002, p. 8532-8539, Vol. 76, No. 17
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.17.8532-8539.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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