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Related Experiment Videos

Drosophila ARSs contain the yeast ARS consensus sequence and a replication enhancer.

J S Mills, A J Kingsman, S M Kingsman

    Nucleic Acids Research
    |August 26, 1986
    PubMed
    Summary

    Researchers identified complex autonomously replicating sequences (ARSs) in fruit fly DNA that function in yeast. These fruit fly ARS elements have distinct replication and enhancer domains, suggesting conserved mechanisms for DNA replication across species.

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    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Autonomously replicating sequences (ARSs) are crucial for DNA replication initiation in eukaryotic organisms.
    • Understanding ARS function in different species can reveal conserved mechanisms of DNA replication.

    Purpose of the Study:

    • To isolate and characterize autonomously replicating sequences (ARSs) from Drosophila melanogaster (fruit fly) DNA.
    • To compare the functional behavior of Drosophila ARSs in yeast with known yeast ARS elements.
    • To elucidate the structural and functional domains of Drosophila ARSs.

    Main Methods:

    • Isolation of restriction fragments from Drosophila melanogaster DNA.
    • Functional analysis of cloned DNA fragments in yeast using plasmids.

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  • Subcloning and BAL-31 deletion analysis to localize ARS functions.
  • Sequence analysis to identify conserved motifs.

    • Main Results:

    • Several DNA fragments from Drosophila melanogaster exhibited ARS activity in yeast.
    • Drosophila ARS elements were found to be structurally complex, each containing at least two functional domains: a replication sequence (RS) and a replication enhancer (RE).
    • The RS domain of three characterized Drosophila ARSs contained an 11 bp sequence identical to the yeast ARS consensus sequence.

    Conclusions:

    • Drosophila melanogaster ARSs possess a complex structure with distinct functional domains essential for replication and enhancer activity.
    • The presence of a conserved yeast ARS consensus sequence in Drosophila ARSs suggests potential evolutionary conservation of DNA replication mechanisms.
    • These findings support the hypothesis that heterologous ARS elements can be biologically significant and functionally conserved across species.