The fate of the initiator tRNAs is sensitive to the critical balance between interacting proteins.
Formylation of the initiator tRNA is essential for normal growth of Escherichia coli. The initiator tRNA containing the U35A36 mutation (CUA anticodon) initiates from UAG codon. However, an additional mutation at position 72 (72A --> G) renders the tRNA (G72/U35A36) inactive in initiation because it is defective in formylation. In this study, we isolated U1G72/U35A36 tRNA containing a wobble base pair at 1-72 positions as an intragenic suppressor of the G72 mutation. The U1G72/U35A36 tRNA is formylated and participates in initiation. More importantly, we show that the mismatch at 1-72 positions of the initiator tRNA, which was thus far thought to be the hallmark of the resistance of this tRNA against peptidyl-tRNA hydrolase (PTH), is not sufficient. The amino acid attached to the initiator tRNA is also important in conferring protection against PTH. Further, we show that the relative levels of PTH and IF2 influence the path adopted by the initiator tRNAs in protein synthesis. These findings provide an important clue to understand the dual function of the single tRNA(Met) in initiation and elongation, in the mitochondria of various organisms.
Thanedar
S
S
Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560 012, India.
Kumar
N V
NV
Varshney
U
U
D000926:Anticodon; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D003062:Codon; D018387:Codon, Initiator; D004926:Escherichia coli / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation; D010442:Peptide Chain Initiation, Translational / Q000235:genetics*; D010448:Peptide Initiation Factors / Q000378:metabolism; D010957:Plasmids; D039881:Prokaryotic Initiation Factor-2; D012329:RNA, Bacterial / Q000235:genetics; D012343:RNA, Transfer / Q000235:genetics*; D012358:RNA, Transfer, Met / Q000235:genetics
D000926:Anticodon; D003062:Codon; D018387:Codon, Initiator; D010448:Peptide Initiation Factors; D039881:Prokaryotic Initiation Factor-2; D012329:RNA, Bacterial; D012358:RNA, Transfer, Met; C024988:tRNA, formylmethionine-; D012343:RNA, Transfer; D002265:Carboxylic Ester Hydrolases; C019674:aminoacyl-tRNA hydrolase