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Related Experiment Videos

Bent DNA at a yeast autonomously replicating sequence.

M Snyder, A R Buchman, R W Davis

    Nature
    |November 6, 1986
    PubMed
    Summary
    This summary is machine-generated.

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    Bent DNA, characterized by specific poly(A) stretches, was identified at a yeast replication origin (ARS1). This bent DNA region binds a protein factor and is crucial for ARS function.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Bent DNA, DNA exhibiting abnormal electrophoretic mobility, is found in prokaryotes and eukaryotes.
    • Previous studies identified bent DNA at replication origins in lambda phage and simian virus 40 (SV40).
    • Kinetoplast DNA has also shown evidence of being curved or 'bent'.

    Purpose of the Study:

    • To investigate the presence and characteristics of bent DNA at a yeast autonomously replicating sequence (ARS1).
    • To determine the functional significance of bent DNA at the ARS1 replication origin.
    • To identify potential DNA-binding proteins associated with the bent DNA region.

    Main Methods:

    • Electrophoretic mobility analysis to detect bent DNA.
    • DNA sequencing and structural analysis to characterize the bent DNA segment.

    Related Experiment Videos

  • In vivo functional assays to assess the impact of bent DNA on ARS function.
  • Chromatin immunoprecipitation or similar techniques to identify DNA-binding proteins.

    • Main Results:

    • Bent DNA was localized to a 40-55 base pair (bp) segment within the yeast ARS1 region.
    • This segment contains six poly(A) stretches (3-5 nucleotides long) phased approximately every 10.5 bp.
    • A DNA-binding site for a yeast protein factor was identified within the bent DNA region.
    • This region is located at the 3' end of the TRP1 gene, nucleosome-free, and 80 bp from the ARS consensus sequence.
    • Deletion of this bent DNA region impaired ARS function in vivo.

    Conclusions:

    • Bent DNA is present at the yeast ARS1 replication origin and is characterized by specific poly(A) repeat phasing.
    • The bent DNA region serves as a binding site for a yeast protein factor, suggesting regulatory roles.
    • This bent DNA element is essential for efficient ARS function, potentially influencing transcription termination or nucleosome assembly.