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Nuclear morphology during the S phase

G Mazzotti1, P Gobbi, L Manzoli

  • 1Istituto Anatomia Umana Normale, Università di Bologna, Italy.

Microscopy Research and Technique
|April 4, 1998
PubMed
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This study visualizes DNA replication during the S phase using Bromodeoxyuridine (BrdU) labeling. Findings suggest DNA replication occurs on a nucleosomal fiber level with enzymes moving along a fixed DNA template.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Chromatin Structure

Background:

  • Understanding chromatin organization during DNA replication is crucial for cell cycle regulation.
  • Previous studies have provided insights into DNA replication but lacked ultrastructural detail of chromatin dynamics.

Purpose of the Study:

  • To investigate the ultrastructural arrangement of chromatin during the S phase of the cell cycle.
  • To determine the localization and organization of DNA replication units within the nucleus.

Main Methods:

  • Immunogold detection of Bromodeoxyuridine (BrdU) in synchronized cells (3T3 fibroblasts, regenerating liver, Friend Leukemia Cells).
  • Cell synchronization and BrdU pulse-labeling.
  • Scanning transmission electron microscopy (STEM) and computer image analysis.

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Main Results:

  • BrdU labeling revealed 10-nm fibers as replication units, organized in a lacework pattern.
  • DNA replication sites shift from dispersed chromatin in early S phase to perinuclear heterochromatin in late S phase.
  • Dispersed chromatin exhibits a regular, ring-like arrangement of 10-nm fibers, confirmed by BrdU distribution.

Conclusions:

  • DNA replication occurs at the level of nucleosomal-like fibers.
  • The study proposes a model where replicating enzyme machinery moves over a fixed DNA template.
  • Chromatin condensation into heterochromatin can occur post-replication without translocation.