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Related Experiment Videos

DNA cruciforms and the nuclear supporting structure.

G K Ward1, A Shihab-el-Deen, M Zannis-Hadjopoulos

  • 1McGill Cancer Centre, McGill University, Montreal, Quebec, Canada.

Experimental Cell Research
|July 1, 1991
PubMed
Summary
This summary is machine-generated.

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DNA cruciforms, potential replication origins, are mostly digested by DNase I, suggesting they are not tightly bound to nuclear matrix proteins. Some resistant cruciforms associate with nuclear membrane and nucleolus, and none are found in metaphase chromosomes.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • DNA cruciforms are hypothesized as recognition sites at eukaryotic DNA replication origins.
  • Monoclonal antibodies specific to DNA cruciforms have been generated.
  • Previous studies showed these antibodies increase DNA synthesis and gene copy number, suggesting cruciform stabilization at replication origins.

Purpose of the Study:

  • To investigate the association of DNA cruciforms with the nuclear matrix and chromosomal scaffold.
  • To understand the in vivo localization and dynamics of cruciform structures.

Main Methods:

  • Utilized DNase I digestion to assess the accessibility of cruciforms.
  • Employed fluorescent labeling with anti-cruciform antibodies to visualize cruciform distribution in cells.

Related Experiment Videos

  • Examined cruciform presence in metaphase chromosomes.

    • Main Results:

    • Most cruciforms detected in G1/S phase cells are susceptible to DNase I digestion, indicating limited association with nuclear matrix proteins.
    • A DNase I-resistant fraction of cruciforms was observed to be associated with the nuclear membrane and nucleolus.
    • Cruciforms were undetectable in metaphase chromosomes, suggesting they are transient or inaccessible.

    Conclusions:

    • DNA cruciforms may not be tightly bound to nuclear matrix proteins during most of the cell cycle.
    • The transient nature of cruciforms is supported by their absence in metaphase chromosomes.
    • Cruciforms likely interact with proteins involved in replication or transcription.