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Journal of Virology, April 2000, p. 3141-3148, Vol. 74, No. 7
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Complex Effects of Deletions in the 5' Untranslated Region of Primate Foamy Virus on Viral Gene Expression and RNA Packaging

Martin Heinkelein,1 Jana Thurow,1 Marco Dressler,1 Horst Imrich,1 Dieter Neumann-Haefelin,2 Myra O. McClure,3 and Axel Rethwilm1,4,*

Institut für Virologie und Immunbiologie, Universität Würzburg, Wurzburg,1 Abteilung für Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, Freiburg,2 and Institut für Virologie, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Dresden,4 Germany, and Jefferiss Research Trust Laboratories, Imperial College of Medicine at St. Mary's, London, United Kingdom3

Received 16 September 1999/Accepted 27 December 1999

Due to various advantageous features there is current interest in retroviral vectors derived from primate foamy viruses (PFVs). Two PFV cis-acting sequences have been mapped in the 5' region of the RNA (pre-)genome and in the 3' pol genomic region. In order to genetically separate PFV packaging constructs from vector constructs, we investigated the effect of deletions in the 5' untranslated region (UTR) of PFV packaging constructs and vectors on gene expression and RNA incorporation into viral particles. Our results indicate that the 5' UTR serves different previously unknown functions. First, the R region of the long terminal repeat was found to be required for PFV gag gene expression. This regulation of gene expression appeared to be mainly posttranscriptional. Second, constructs with sequence deletions between the R region and the gag gene start codon packaged as much viral mRNA into particles as the undeleted construct, and RNA from such a 5'-UTR-deleted packaging construct was copackaged into vector-virus particles, together with vector RNA which was preferentialy packaged. Finally, in the U5 region a sequence was identified that was required to allow cleavage of the Gag precursor protein by the pol gene-encoded protease, suggesting a role of RNA in PFV particle formation. Taken together, the results indicate that complex interactions of the viral RNA, capsid, and polymerase proteins take place during PFV particle formation and that a clear separation of PFV vector and packaging construct sequences may be difficult to achieve.

* Corresponding author. Mailing address: Institut für Virologie, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Gerichtsstr. 5, 01069 Dresden, Germany. Phone: (49) 351-441-5739. Fax: (49) 351-459-3530. E-mail: Axel.Rethwilm{at}mailbox.tu-dresden.de.

Journal of Virology, April 2000, p. 3141-3148, Vol. 74, No. 7
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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