A reduced level of charged tRNAArgmnm5UCU triggers the wild-type peptidyl-tRNA to frameshift.
Frameshift mutations can be suppressed by a variety of differently acting external suppressors. The +1 frameshift mutation hisC3072, which has an extra G in a run of Gs, is corrected by the external suppressor mutation sufF44. We have shown that sufF44 and five additional allelic suppressor mutations are located in the gene argU coding for the minor tRNAArgmnm5UCU and alter the secondary and/or tertiary structure of this tRNA. The C61U, G53A, and C32U mutations influence the stability, whereas the C56U, C61U, G53A, and G39A mutations decrease the arginylation of tRNAArgmnm5UCU. The T-10C mutant has a base substitution in the -10 consensus sequence of the argU promoter that reduces threefold the synthesis of tRNAArgmnm5UCU . The lower amount of tRNAArgmnm5UCU or impaired arginylation, either independently or in conjunction, results in inefficient reading of the cognate AGA codon that, in turn, induces frameshifts. According to the sequence of the peptide produced from the suppressed -GGG-GAA-AGA- frameshift site, the frameshifting tRNA in the argU mutants is tRNAGlumnm5s2UUC, which decodes the GAA codon located upstream of the AGA arginine codon, and not the mutated tRNAArgmnm5UCU. We propose that an inefficient decoding of the AGA codon by a defective tRNAArgmnm5UCU stalls the ribosome at the A-site codon allowing the wild-type form of peptidyl-tRNAGlumnm5s2UUC to slip forward 1 nucleotide and thereby re-establish the ribosome in the 0-frame. Similar frame-shifting events could be the main cause of various phenotypes associated with environmental or genetically induced changes in the levels of aminoacylated tRNA.
Leipuviene
Ramune
R
Department of Molecular Biology, Umeå University, S-90187 Umeå, Sweden.
Björk
Glenn R
GR
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D003062:Codon / Q000235:genetics; D016368:Frameshift Mutation / Q000235:genetics; D018965:Frameshifting, Ribosomal / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D008957:Models, Genetic; D008969:Molecular Sequence Data; D012347:RNA, Transfer, Arg / Q000235:genetics* / Q000378:metabolism*; D012352:RNA, Transfer, Glu / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D019779:Salmonella enterica / Q000235:genetics*; D013489:Suppression, Genetic / Q000235:genetics; D046249:Transfer RNA Aminoacylation
D003062:Codon; D012347:RNA, Transfer, Arg; D012352:RNA, Transfer, Glu; D011993:Recombinant Fusion Proteins