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GST-MNV VPg 102–124 inhibits MNV VPg-mediated translation in vitro but not cap-dependent or IRES-dependent translation.

figure

posted on 2016-01-18, 14:36 authored by Eoin N. Leen, Frédéric Sorgeloos, Samantha Correia, Yasmin Chaudhry, Fabien Cannac, Chiara Pastore, Yingqi Xu, Stephen C. Graham, Stephen J. Matthews, Ian G. Goodfellow, Stephen Curry

In vitro translation reactions programmed with VPg-linked MNV RNA were performed in the presence of increasing concentrations of GST-MNV VPg(102–124) WT protein or the GST-MNV VPg(102–124) F123A mutant that binds very poorly to eIF4G. Protein synthesis was monitored by autoradiography of SDS PAGE analysis of incorporation of ³⁵S-methionine in translation reactions. (A) Top panel: Effect of exogenous GST-MNV VPg(102–124) proteins on translation from VPg-linked MNV RNA; bottom panel: quantitative analysis of the level of ³⁵S-methionine incorporation. This result was confirmed by independent replication–see S5A Fig. (B) Top panel: Effect of exogenous GST-MNV VPg(102–124) proteins on translation from capped bi-cistronic mRNA constructs containing the FMDV IRES between the first (CAT) and second (Luc) cistrons; bottom panel: quantitative analysis of the level of ³⁵S-methionine incorporation.