Roles of yeast eIF2α and eIF2β subunits in the binding of the initiator methionyl-tRNA

Marie Naveau (1) , Christine Lazennec-Schurdevin (1) , Michel Panvert (1) , Etienne Dubiez (1) , Yves Mechulam (1) , Emmanuelle Schmitt (1)

1 BIOC - Laboratoire de Biochimie de l'Ecole polytechnique

Heterotrimeric eukaryotic/archaeal translation initiation factor 2 (e/aIF2) binds initiator methionyl-tRNA and plays a key role in the selection of the start codon on messenger RNA. tRNA binding was extensively studied in the archaeal system. The γ subunit is able to bind tRNA, but the α subunit is required to reach high affinity whereas the β subunit has only a minor role. In Saccharomyces cerevisiae however, the available data suggest an opposite scenario with β having the most important contribution to tRNA-binding affinity. In order to overcome difficulties with purification of the yeast eIF2γ subunit, we designed chimeric eIF2 by assembling yeast α and β subunits to archaeal γ subunit. We show that the β subunit of yeast has indeed an important role, with the eukaryote-specific N- and C-terminal domains being necessary to obtain full tRNA-binding affinity. The α subunit apparently has a modest contribution. However, the positive effect of α on tRNA binding can be progressively increased upon shortening the acidic C-terminal extension. These results, together with small angle X-ray scattering experiments, support the idea that in yeast eIF2, the tRNA molecule is bound by the α subunit in a manner similar to that observed in the archaeal aIF2-GDPNP-tRNA complex. © The Author(s) 2012. Published by Oxford University Press.

Domains: Life Sciences [q-bio]; Biochemistry, Molecular Biology

Marie Naveau, Christine Lazennec-Schurdevin, Michel Panvert, Etienne Dubiez, Yves Mechulam, et al.. Roles of yeast eIF2α and eIF2β subunits in the binding of the initiator methionyl-tRNA. Nucleic Acids Research, 2013, 41 (1), pp.1047-1057. ⟨10.1093/nar/gks1180⟩. ⟨hal-00840386⟩