Junín virus Z protein is packaged into exportable vesicles after expression in mammalian cells

BORIO, CS; BILEN, MF; URE, A; STEPHAN, BI; ISERTE, JA; GOÑI, SE; GOMEZ, R; GHIRINGHELLI, PD; LOZANO, ME

Tucumán

Congreso; SAIB; 2009

SAIB

Junín virus, the etiological agent of Argentine hemorrhagic fever, belongs to the Arenaviridae family. This family has 23 recognized species with an ambisense coding-bipartite ssRNA genome. The enveloped virions include non-equimolar amounts of each genomic RNA, named L (ca. 7200 nt; coding for L and Z proteins), and S (ca. 3500 nt; coding for N and GPC proteins). The Z protein, the matrix protein of the virion, interacts with several proteins in infected cells. The N-terminal region has a conserved G at position 2, which would be myristoilated, and the C-terminal region comprises the late domain, previously described as the main driving force of the budding process. In this study we analyzed the Z protein expression and its ability to induce exported vesicle formation. We used a modified Z protein in which the C-terminal aminoacid is fused to GFP protein (Z-GFP), and analyzed its expression and exportation potential in mammal cells by differential ultracentrifugation followed by western blotting, flow citometry, fluorescent microscopy and immunogold-electron microscopy. Our results proved that Z-GFP protein expression induce virus like particles formation which contains the recombinant protein.