Translational roles of the C75 2'OH in an in vitro tRNA transcript at the ribosomal A, P and E sites.

ABSTRACT: Aminoacyl-tRNAs containing a deoxy substitution in the penultimate nucleotide (C75 2'OH → 2'H) have been widely used in translation for incorporation of unnatural amino acids (AAs). However, this supposedly innocuous modification surprisingly increased peptidyl-tRNA^Ala_ugc drop off in biochemical assays of successive incorporations. Here we predict the function of this tRNA 2'OH in the ribosomal A, P and E sites using recent co-crystal structures of ribosomes and tRNA substrates and test these structure-function models by systematic kinetics analyses. Unexpectedly, the C75 2'H did not affect A- to P-site translocation nor peptidyl donor activity of tRNA^Ala_ugc. Rather, the peptidyl acceptor activity of the A-site Ala-tRNA^Ala_ugc

SUBMITTER: Wang J 

PROVIDER: S-EPMC5532260 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature