Author: Madan, Vanesa; Redondo, Natalia; Carrasco, Luis
Title: Cell permeabilization by poliovirus 2B viroporin triggers bystander permeabilization in neighbouring cells through a mechanism involving gap junctions Cord-id: mxcoztak Document date: 2010_3_12
ID: mxcoztak
Snippet: Poliovirus 2B protein is a wellâ€known viroporin implicated in plasma membrane permeabilization to ions and lowâ€molecularâ€weight compounds during infection. Translation in mammalian cells expressing 2B protein is inhibited by hygromycin B (HB) but remains unaffected in mock cells, which are not permeable to the inhibitor. Here we describe a previously unreported bystander effect in which healthy baby hamster kidney (BHK) cells become sensitive to HB when coâ€cultured with a low proportion
Document: Poliovirus 2B protein is a wellâ€known viroporin implicated in plasma membrane permeabilization to ions and lowâ€molecularâ€weight compounds during infection. Translation in mammalian cells expressing 2B protein is inhibited by hygromycin B (HB) but remains unaffected in mock cells, which are not permeable to the inhibitor. Here we describe a previously unreported bystander effect in which healthy baby hamster kidney (BHK) cells become sensitive to HB when coâ€cultured with a low proportion of cells expressing poliovirus 2B. Viroporins E from mouse hepatitis virus, 6K from Sindbis virus and NS4A protein from hepatitis C virus were also able to permeabilize neighbouring cells to different extents. Expression of 2B induced permeabilization of neighbouring cell lines other than BHK. We found that gap junctions are responsible mediating the observed bystander permeabilization. Gap junctional communication was confirmed in 2Bâ€expressing coâ€cultures by fluorescent dye transfer. Moreover, the presence of connexin 43 was confirmed in both mock and 2Bâ€transfected cells. Finally, inhibition of HB entry to neighbouring cells was observed with 18αâ€glycyrrhethinic acid, an inhibitor of gap junctions. Taken together, these findings support a mechanism involving gap junctional intercellular communication in the bystander permeabilization effect observed in healthy cells coâ€cultured with poliovirus 2Bâ€expressing cells.
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