Capsid-deficient alphaviruses generate propagative infectious microvesicles at the plasma membrane
Ruiz-Guillen M (1,2,3), Gabev E (1,2,4), Quetglas JI (1,2), Casales E (1,2), Ballesteros-Briones MC (1,2), Poutou J (1,2), Aranda A (1,2,5), Martisova E (1,2), Bezunartea J (1,2,6), Ondiviela M (7), Prieto J (1,2,8), Hernandez-Alcoceba R (1,2), Abrescia NG (7,9), Smerdou C (10,11).
(1) Division of Gene Therapy, CIMA, University of Navarra, Pamplona, Spain.
(2) IdiSNA, Navarra Institute for Health Research, Pamplona, Spain.
(3) 3P Biopharmaceuticals S.L., Noain, Spain.
(4) Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy.
(5) UFR des Sciences de la Santé Simone Veil, 2 avenue de la Source de la Bievre, 78180, Montugny-le-Bretonneux, France.
(6) Experimental Ophthalmology Laboratory, School of Medicine, University of Navarra, Pamplona, Spain.
(7) Structural Biology Unit, CIC bioGUNE, CIBERehd, Bizkaia Technology Park, Derio, Spain.
(8) Liver Unit, Clinica Universitaria de Navarra, CIBERehd, Pamplona, Spain.
(9) IKERBASQUE, Basque Foundation for Science, Bilbao, Spain.
(10) Division of Gene Therapy, CIMA, University of Navarra, Pamplona, Spain.
(11) IdiSNA, Navarra Institute for Health Research, Pamplona, Spain.
Alphavirus budding is driven by interactions between nucleocapsids assembled in the cytoplasm and envelope proteins present at the plasma membrane. So far, the expression of capsid and envelope proteins in infected cells has been considered an absolute requirement for alphavirus budding and propagation.
In the present study, we show that Semliki Forest virus and Sindbis virus lacking the capsid gene can propagate in mammalian and insect cells. This propagation is mediated by the release of infectious microvesicles (iMVs), which are pleomorphic and have a larger size and density than wild-type virus. iMVs, which contain viral RNA inside and viral envelope proteins on their surface, are released at the plasma membrane and infect cells using the endocytic pathway in a similar way to wild-type virus.
iMVs are not pathogenic in immunocompetent mice when injected intravenously, but can infect different organs like lungs and heart. Finally, we also show that alphavirus genomes without capsid can mediate the propagation of heterologous genes, making these vectors potentially interesting for gene therapy or vaccination studies.
The minimalist infectious system described in this study shows that a self-replicating RNA able to express membrane proteins with binding and fusion properties is able to propagate, providing some insights into virus evolution.
CITATION Cell Mol Life Sci. 2016 Apr 27