Abstract

BackgroundThe mRNA translation initiation region (TIR) comprises the initiator codon, Shine-Dalgarno (SD) sequence and translational enhancers. Probably the most abundant class of enhancers contains A/U-rich sequences. We have tested the influence of SD sequence length and the presence of enhancers on the efficiency of translation initiation.ResultsWe found that during bacterial growth at 37°C, a six-nucleotide SD (AGGAGG) is more efficient than shorter or longer sequences. The A/U-rich enhancer contributes strongly to the efficiency of initiation, having the greatest stimulatory effect in the exponential growth phase of the bacteria. The SD sequences and the A/U-rich enhancer stimulate translation co-operatively: strong SDs are stimulated by the enhancer much more than weak SDs. The bacterial growth rate does not have a major influence on the TIR selection pattern. On the other hand, temperature affects the TIR preference pattern: shorter SD sequences are preferred at lower growth temperatures. We also performed an in silico analysis of the TIRs in all E. coli mRNAs. The base pairing potential of the SD sequences does not correlate with the codon adaptation index, which is used as an estimate of gene expression level.ConclusionIn E. coli the SD selection preferences are influenced by the growth temperature and not influenced by the growth rate. The A/U rich enhancers stimulate translation considerably by acting co-operatively with the SD sequences.

Highlights

  • The mRNA translation initiation region (TIR) comprises the initiator codon, ShineDalgarno (SD) sequence and translational enhancers

  • The 30S subunit complexed with IF1 and IF3 binds to the mRNA, followed by IF2 and GTP-dependent binding of fMet-tRNAfMet [2]

  • The mRNAs synthesized from the tac promoter contained a lacO operator sequence in front of the TIRs

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Summary

Introduction

The mRNA translation initiation region (TIR) comprises the initiator codon, ShineDalgarno (SD) sequence and translational enhancers. The 30S ribosomal subunit, assisted by initiation factors (IF) 1, 2 and 3 and fMet-tRNAfMet, recognizes the translation initiation region (TIR) of the mRNA. This event is followed by binding of the 50S ribosomal subunit and release of the initiation factors [1]. The 30S subunit complexed with IF1 and IF3 binds to the mRNA, followed by IF2 and GTP-dependent binding of fMet-tRNAfMet [2]. The IF2:GTP:fMet-tRNAfMet complex binds to the 30S subunit followed by mRNA recognition [3]. The relative frequencies with which these pathways are used in bacterial cells are currently not clear

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