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Artificial mitochondrial presequences.

D S Allison, G Schatz

    Proceedings of the National Academy of Sciences of the United States of America
    |December 1, 1986
    PubMed
    Summary
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    Artificial mitochondrial presequences, composed of specific amino acids like arginine, serine, and leucine, successfully targeted proteins to mitochondria. This suggests the balance of amino acid types, not specific sequences, is key for mitochondrial targeting.

    Area of Science:

    • Mitochondrial biology
    • Protein targeting
    • Molecular genetics

    Background:

    • Mitochondrial presequences are essential for directing proteins to mitochondria.
    • The precise amino acid composition and sequence requirements for presequence function are not fully understood.

    Purpose of the Study:

    • To investigate whether specific amino acid sequences or the overall balance of amino acid types in mitochondrial presequences dictates their targeting function.
    • To construct and test artificial presequences with defined amino acid compositions.

    Main Methods:

    • Synthetic oligonucleotides were used to create artificial mitochondrial presequences.
    • These artificial presequences, containing specific ratios of arginine, serine, and leucine, were fused to yeast cytochrome oxidase subunit IV.

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  • The functionality of these constructs was assessed by observing protein localization in vivo and import into isolated yeast mitochondria.

    • Main Results:

    • Artificial presequences composed of arginine, serine, and leucine successfully directed cytochrome oxidase subunit IV to the mitochondria.
    • These sequences mediated the import of subunit IV into isolated yeast mitochondria.
    • Artificial sequences with glutamine, arginine, and serine were inactive, indicating the importance of specific amino acid properties.

    Conclusions:

    • Mitochondrial presequence targeting is not dependent on specific amino acid sequences.
    • The overall balance of basic, hydrophobic, and hydroxylated amino acids is crucial for mitochondrial presequence function.
    • This finding provides new insights into the principles of protein import into mitochondria.