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Sequences essential for IS50 transposition. The first base-pair.

C Sasakawa, S H Phadnis, G F Carle

    Journal of Molecular Biology
    |April 20, 1985
    PubMed
    Summary
    This summary is machine-generated.

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    The first base pair at each end of the insertion sequence IS50 is crucial for transposition. Mutations at this site significantly decrease transposition frequency, highlighting its importance for transposase binding and function.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • Insertion sequences (IS) are mobile genetic elements that can move within a genome.
    • The transposition of IS50, mediated by its encoded transposase, is essential for its mobility.
    • Specific DNA sequences at the ends of IS elements typically serve as recognition sites for the transposase.

    Purpose of the Study:

    • To investigate the role of the terminal base pair at each end of the insertion sequence IS50 in its transposition process.
    • To determine if the very first base pair of IS50 is essential for transposase recognition and activity.

    Main Methods:

    • Site-directed mutagenesis was used to introduce specific base-pair changes (5'C to 5'G transversions) at the 5' ends of IS50.
    • Transposition frequency was quantified for the wild-type and mutant IS50 elements.

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  • DNA sequence analysis was performed to confirm the presence of the mutation in the transposed products.

    • Main Results:

    • Mutations changing the first base pair at each end of IS50 to guanine drastically reduced transposition frequency to 1-2% of wild-type levels.
    • DNA sequence analysis confirmed that the introduced 5'G mutation was maintained in the transposed DNA fragments.

    Conclusions:

    • The terminal base pair at each end of IS50 is essential for efficient transposition.
    • These findings suggest that the first base pair is critical for transposase binding or catalytic activity during transposition.