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Replication pattern of human repeated DNA sequences.

R Meneveri, A Agresti, D Breviario

    Biochimica Et Biophysica Acta
    |October 5, 1984
    PubMed
    Summary
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    Human repetitive DNA sequences, including alpha RI-DNA and alpha satellite sequences, replicate throughout the S phase. DNA replication patterns show variations in fragment intensity during different S periods, with a specific XbaI fragment becoming undetectable in late S.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Cell Biology

    Background:

    • Understanding DNA replication timing is crucial for comprehending genome stability and gene regulation.
    • Human repetitive DNA families, such as alpha RI-DNA and alpha satellite sequences, constitute significant portions of the genome and their replication dynamics are not fully elucidated.

    Purpose of the Study:

    • To investigate the replication patterns of specific human repetitive DNA sequences (alpha RI-DNA and alpha satellite sequences) during different phases of the S period.
    • To analyze the DNA replication timing of these sequences in synchronized HL-60 cells.

    Main Methods:

    • Cell synchronization using aphidicolin or thymidine.
    • Labeling of newly synthesized DNA with bromodeoxyuridine (BUdR) or [3H]thymidine during distinct S phase intervals.

    Related Experiment Videos

  • Hybridization techniques to detect alpha RI-DNA replication.
  • Analysis of XbaI and HaeIII digested DNA fragments to study alpha satellite sequence replication patterns via fluorography and densitometry.

    • Main Results:

    • The EcoRI 340 bp repetitive DNA family (alpha RI-DNA) is replicated throughout the entire S period.
    • DNA synthesized during different S phase periods exhibits a similar ladder of fragments after XbaI and HaeIII digestion, indicating conserved structural organization.
    • The intensity of specific DNA fragments varies across the S phase, with the XbaI 2.4 kb fragment becoming undetectable in late S, suggesting differential replication timing or accessibility.

    Conclusions:

    • Human repetitive DNA sequences, including alpha RI-DNA and alpha satellite sequences, are replicated throughout the S phase.
    • Replication timing of repetitive DNA elements can be characterized by variations in fragment intensity, providing insights into their dynamic behavior during S phase.
    • The findings contribute to a better understanding of the replication dynamics of repetitive DNA in the human genome.