ms2i6A deficiency enhances proofreading in translation.
The hypermodified base 2-methylthio-N6-isopentenyladenosine (ms2i6A) at position 37 occurs frequently in tRNAs that read codons starting with uridine. Here we have studied how ms2i6A affects the accuracy of poly(U) translation in vitro. Deficiency leads to a higher rejection rate of tRNA4(Leu) by more aggressive proofreading on the wild-type ribosome, but with the initial selection step unchanged. Our data indicate that ms2i6A has no effect on codon-anticodon interactions on wild-type ribosomes as long as aminoacyl-tRNA is in ternary complex with EF-Tu and GTP. ms2i6A deficiency in the cognate poly(U) reader tRNA(Phe) leads to increased misreading when the near-cognate competitor tRNA4(Leu) is wild-type. ms2i6A deficiency in tRNA4(Leu) gives a decreased error level in competition with wild-type tRNA(Phe).
Díaz
I
I
Department of Molecular Biology, University of Uppsala, Sweden.
Ehrenberg
M
M
D003062:Codon; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D007541:Isopentenyladenosine / Q000031:analogs & derivatives* / Q000235:genetics; D007700:Kinetics; D011072:Poly U; D014176:Protein Biosynthesis; D012356:RNA, Transfer, Leu / Q000235:genetics*; D012360:RNA, Transfer, Phe / Q000235:genetics*; D012270:Ribosomes / Q000378:metabolism*
D003062:Codon; D012356:RNA, Transfer, Leu; D012360:RNA, Transfer, Phe; C008503:2-methylthio-N-6-isopentenyladenosine; D011072:Poly U; D007541:Isopentenyladenosine; D006160:Guanosine Triphosphate