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Related Experiment Videos

Selection of AUG initiation codons differs in plants and animals.

H A Lütcke, K C Chow, F S Mickel

    The EMBO Journal
    |January 1, 1987
    PubMed
    Summary

    Nucleotide sequences near the start codon influence protein synthesis initiation. Differences in translation efficiency were observed in reticulocyte lysate but not wheat germ systems, suggesting distinct initiation mechanisms in plants and animals.

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    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Protein synthesis initiation is a critical regulatory step in gene expression.
    • The sequence context surrounding the AUG start codon plays a role in translation initiation efficiency.

    Purpose of the Study:

    • To investigate the impact of the nucleotide at position -3 relative to the AUG start codon on protein synthesis initiation.
    • To compare translation initiation mechanisms in different in vitro systems and across species.

    Main Methods:

    • Synthetic messenger RNAs (mRNAs) with variations at nucleotide -3 were created using SP6 promoter-directed cDNA transcription.
    • In vitro translation assays were performed using reticulocyte lysate and wheat germ systems.
    • Analysis of plant and animal mRNA sequences to determine consensus sequences around AUG initiation codons.

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    Main Results:

    • A hierarchy of translational efficiency was observed in the reticulocyte lysate system, with nucleotide A at position -3 yielding the highest efficiency (100%), followed by G (85%), U (61%), and C (38%).
    • No significant differences in translational efficiency were observed in the wheat germ system.
    • Differential mRNA degradation or polypeptide chain elongation were ruled out as explanations for the observed differences in the reticulocyte lysate.
    • mRNA competition assays exacerbated efficiency differences in reticulocyte lysate but had no effect in wheat germ.
    • Consensus sequences around AUG initiation codons differed between plants (AACAAUGGC) and animals (CACCAUG).

    Conclusions:

    • The nucleotide at position -3 significantly influences translation initiation efficiency in a reticulocyte lysate system.
    • Plant and animal translation initiation factors exhibit distinct preferences for start codon context, suggesting divergent mechanisms for selecting AUG initiation codons.