Efficient Cleavage of p220 by Poliovirus 2Apro Expression in Mammalian Cells: Effects on Vaccina Virus

Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease

“…Mouse monoclonal antibodies were used to detect nucleolin (MS3 (26) and GFP (Clontech, catalog no. 632381), whereas rabbit polyclonal antiserum was used to detect eIF4G (27) and NF-B (Santa Cruz Biotechnology, Inc., catalog no. sc-109).…”

RNA nuclear export is blocked by poliovirus 2A protease and is concomitant with nucleoporin cleavage

“…Subcellular fractions were obtained using a buffer containing NP-40, RNaseOUT (Invitrogen) and a cocktail of protease inhibitors (Sigma-Aldrich) as described previously (Perales et al, 2003). eIF4GI was detected with antisera raised against peptides derived from the N-terminal and Cterminal, at 1:1000 dilution (Aldabe et al, 1995). Rabbit antisera against N-terminal and C-terminal region of eIF4GII (a generous gift from Nahum Sonenberg, McGill University, Montreal, Canada) were employed at 1:500 dilution.…”

A Stable HeLa Cell Line That Inducibly Expresses Poliovirus 2A pro : Effects on Cellular and Viral Gene Expression

“…The recombinant vaccinia virus bearing the T7 RNA polymerase (VT7) (20) was kindly provided by B. Moss (National Institutes of Health, Bethesda, Md.). Infection of HeLa cells with vaccinia virus and VT7 has been previously described (1,3).…”

“…Attempts to express 2A pro from recombinant vaccinia viruses have been hampered by 2A pro toxicity (30,62). Transient expression of poliovirus 2A pro has been achieved by infection with recombinant vaccinia virus that expresses the T7 RNA polymerase (2,3,19). Poliovirus 2A pro strongly inhibits vaccinia virus gene expression at the translational level (19).…”