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Journal of Virology, April 2007, p. 3198-3205, Vol. 81, No. 7
0022-538X/07/$08.00+0 doi:10.1128/JVI.02655-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.
The Glycoprotein Cytoplasmic Tail of Uukuniemi Virus (Bunyaviridae) Interacts with Ribonucleoproteins and Is Critical for Genome Packaging
Anna K. Överby,
Ralf F. Pettersson,
* and
Etienne P. A. Neve
Ludwig Institute for Cancer Research, Stockholm Branch, Karolinska Institute, Box 240, SE-17177 Stockholm, Sweden
Received 1 December 2006/
Accepted 4 January 2007
We have analyzed the importance of specific amino acids in the cytoplasmic tail of the glycoprotein GN for packaging of ribonucleoproteins (RNPs) into virus-like particles (VLPs) of Uukuniemi virus (UUK virus), a member of the Bunyaviridae family. In order to study packaging, we added the GN/GC glycoprotein precursor (p110) to a polymerase I-driven minigenome rescue system to generate VLPs that are released into the supernatant. These particles can infect new cells, and reporter gene expression can be detected. To determine the role of UUK virus glycoproteins in RNP packaging, we performed an alanine scan of the glycoprotein GN cytoplasmic tail (amino acids 1 to 81). First, we discovered three regions in the tail (amino acids 21 to 25, 46 to 50, and 71 to 81) which are important for minigenome transfer by VLPs. Further mutational analysis identified four amino acids that were important for RNP packaging. These amino acids are essential for the binding of nucleoproteins and RNPs to the glycoprotein without affecting the morphology of the particles. No segment-specific interactions between the RNA and the cytoplasmic tail could be observed. We propose that VLP systems are useful tools for analyzing protein-protein interactions important for packaging of viral genome segments, assembly, and budding of other members of the Bunyaviridae family.
* Corresponding author. Mailing address: Ludwig Institute for Cancer Research, Stockholm Branch, Karolinska Institute, Box 240, SE-17177 Stockholm, Sweden. Phone: 46 (0)8 524 871 01. Fax: 46 (0)8 33 28 12. E-mail:
rpet{at}licr.ki.se.
Published ahead of print on 17 January 2007.
R.F.P. and E.P.A.N. contributed equally to this work.
Journal of Virology, April 2007, p. 3198-3205, Vol. 81, No. 7
0022-538X/07/$08.00+0 doi:10.1128/JVI.02655-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.
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