Ribose 2′-hydroxyl groups in the 5′ strand of the acceptor arm of P-site tRNA are not essential for EF-G catalyzed translocation

Time course of translocation of fragmented tRNA^Met with a single 2′-deoxy substitution in the small fragment. (A) Control time-course experiment showing inhibition of translocation by neomycin. Minus sign (−) indicates absence of EF-G. Neomycin was added to the pretranslocation complex, followed by the addition of EF-G·GTP. Aliquots of the reaction were taken at the indicated time points and analyzed by toeprinting. (B) Translocation of fragmented tRNA^Met incorporating single 2′-deoxy substitution at the indicated positions. EF-G·GTP was added to the pretranslocation complex, and aliquots of the reaction were taken at the indicated time points and mixed with extension reaction mix containing neomycin to stop translocation.