The cooperative function of arginine residues in the Prototype Foamy Virus Gag C-terminus mediates viral and cellular RNA encapsidation

Retrovirology

Dirk Lindemann¹  Winfried Weissenhorn²  Gregory Effantin²  Nicole Stanke¹  Juliane Reh¹  Erik Müllers³  Martin V Hamann¹ 

[1] CRTD/DFG-Center for Regenerative Therapies Dresden - Cluster of Excellence, Technische Universität Dresden, Fetscherstr. 105, Dresden, 01307, Germany;CNRS, UVHCI, Grenoble, F-38000, France;Present address: Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, 171 77, Sweden

关键词: Assembly;    Gag;    RNA packaging;    Foamy virus;   

Others  :  1152283
DOI  :  10.1186/s12977-014-0087-7

 received in 2014-04-16, accepted in 2014-09-23,  发布年份 2014

【 摘 要 】

Background

One unique feature of the foamy virus (FV) capsid protein Gag is the absence of Cys-His motifs, which in orthoretroviruses are irreplaceable for multitude functions including viral RNA genome recognition and packaging. Instead, FV Gag contains glycine-arginine-rich (GR) sequences at its C-terminus. In case of prototype FV (PFV) these are historically grouped in three boxes, which have been shown to play essential functions in genome reverse transcription, virion infectivity and particle morphogenesis. Additional functions for RNA packaging and Pol encapsidation were suggested, but have not been conclusively addressed.

Results

Here we show that released wild type PFV particles, like orthoretroviruses, contain various cellular RNAs in addition to viral genome. Unlike orthoretroviruses, the content of selected cellular RNAs in capsids of PFV vector particles was not altered by viral genome encapsidation. Deletion of individual GR boxes had only minor negative effects (2 to 4-fold) on viral and cellular RNA encapsidation over a wide range of cellular Gag to viral genome ratios examined. Only the concurrent deletion of all three PFV Gag GR boxes, or the substitution of multiple arginine residues residing in the C-terminal GR box region by alanine, abolished both viral and cellular RNA encapsidation (>50 to >3,000-fold reduced), independent of the viral production system used. Consequently, those mutants also lacked detectable amounts of encapsidated Pol and were non-infectious. In contrast, particle release was reduced to a much lower extent (3 to 20-fold).

Conclusions

Taken together, our data provides the first identification of a full-length PFV Gag mutant devoid in genome packaging and the first report of cellular RNA encapsidation into PFV particles. Our results suggest that the cooperative action of C-terminal clustered positively charged residues, present in all FV Gag proteins, is the main viral protein determinant for viral and cellular RNA encapsidation. The viral genome independent efficiency of cellular RNA encapsidation suggests differential packaging mechanisms for both types of RNAs. Finally, this study indicates that analogous to orthoretroviruses, Gag ¿ nucleic acid interactions are required for FV capsid assembly and efficient particle release.

【 授权许可】

   
2014 Hamann et al.; licensee BioMed Central Ltd.

【 图 表 】

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