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The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
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An essential Noc3p dimerization cycle mediates ORC double-hexamer formation in replication licensing.

Aftab Amin1,2, Rentian Wu1, Muhammad Ajmal Khan1

  • 1Division of Life Science, Center for Cancer Research, and State Key Lab of Molecular Neuroscience, Hong Kong University of Science and Technology, Hong Kong, China.

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|January 4, 2023
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A new study reveals how Noc3p protein dimerization regulates DNA replication licensing. This cell cycle-dependent mechanism ensures genome duplication occurs only once per cell cycle, preventing re-replication.

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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Replication licensing ensures DNA replication occurs once per cell cycle.
  • Pre-replicative complexes (pre-RCs) form at replication origins, involving proteins like ORC and Noc3p.
  • ORC dimerization is crucial for MCM double-hexamer loading onto chromatin.

Purpose of the Study:

  • To investigate the role of Noc3p in DNA replication and ribosome biogenesis using Saccharomyces cerevisiae mutants.
  • To elucidate the mechanism by which Noc3p regulates ORC dimerization and pre-RC formation.
  • To understand how Noc3p's cell cycle-dependent dimerization impacts replication licensing.

Main Methods:

  • Utilized separation-of-function NOC3 mutants in Saccharomyces cerevisiae.
  • Analyzed the cell cycle-dependent dimerization of Noc3p and its effect on ORC dimerization.
  • Investigated the role of Noc3p dimerization in replication licensing and origin protection.

Main Results:

  • Confirmed separable functions of Noc3p in DNA replication and ribosome biogenesis.
  • Demonstrated that Noc3p dimerization, occurring at the M-to-G1 transition and reversing in S-phase, regulates ORC dimerization.
  • Showed that coupled Noc3p and ORC dimerization cycles are essential for replication licensing and preventing re-replication.

Conclusions:

  • Noc3p acts as a crucial mediator of ORC dimerization in pre-RC formation.
  • A novel mechanism of replication licensing involving Noc3p and ORC dimerization cycles has been uncovered.
  • This study provides molecular insights into Noc3p's function in cell cycle regulation and genome stability.