Key Points
- To determine the cellular origin and molecular mechanism underlying the formation of poliovirus replication complex vesicles.
- Performed immunofluorescence labeling of viral proteins (2B, 2BC) alongside cellular markers for the endoplasmic reticulum, anterograde transport vesicles, and Golgi apparatus in BT7-H cells.
- Visualized 3D membrane architecture using deconvolved confocal laser scanning microscopy.
- Evaluated vesicle formation following the isolated expression of viral protein 2BC or the P2/P3 polyprotein region.
- Poliovirus vesicles excluded ER-resident markers and colocalized with COPII coat proteins Sec13 and Sec31, confirming the presence of a functional COPII coat and independence from the Golgi complex in early infection.
- Expression of viral protein 2BC or the complete P2 and P3 genomic region independently induced the formation of COPII-coated vesicles morphologically identical to those formed during native infection.
Structured PICO
IInterventionPoliovirus (PV) infection or expression of PV protein 2BC or the entire P2 and P3 genomic region
CComparatorUninfected cells
OOutcomeOrigin and mode of formation of PV vesicles (colocalization of COPII components Sec13 and Sec31)surrogate
Poliovirus replication vesicles are formed at the endoplasmic reticulum utilizing the cellular COPII budding mechanism, homologous to anterograde membrane transport vesicles.