| dc.description.abstract | The enterovirus genus comprises many human pathogens such as poliovirus, coxsackievirus
and rhinovirus. For efficient replication, enteroviruses induce membrane rearrangements
which require the interaction between viral and host factors such as PI4KB, ACBD3 and 3A.
Although interactions between these proteins have been intensively studied, their
localization at replication organelle (RO) membranes is unknown. To investigate components
of ROs in-depth, we established protocols to study coxsackievirus B3 (CVB3) ROs using
expansion microscopy (ExM) and super-resolution microscopy (SRM), i.e. Olympus
SoRaSpin10 system. Our data is the first evidence of 2A localization. The use of SRM allowed
us to visualize the localization of the viral protease 2A at different time points of infection in
greater detail. In addition, we could locate 3A at an early time point after infection (i.e.
3 h p.i.), which was hardly attainable with conventional immunofluorescence microscopy.
Our data suggest that imaging at earlier time points might be possible. Moreover, both viral
proteins were visualized as dotted structures which have not yet been observed. We
demonstrate the potential for in-depth visualization of these viral proteins and their
colocalization with host factors using super-resolution microscopy. The Olympus SoRaSpin10
system offers the possibility to combine SRM with live-cell imaging. Thus, tracking viral
protein localization, RO formation and its dynamics could be further investigated. | |
