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In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
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Before a cell can divide, it must accurately replicate all of its chromosomes, including the DNA and its associated histone and non-histone proteins.  This process begins at numerous origins of replication during the S phase of the cell cycle in each of a cell’s chromosomes simultaneously. Certain nucleotides can act as origins of replication, but these sequences are not well defined - especially in complex, multi-cellular, eukaryotic species. The length of DNA that spans an origin...
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A functional eukaryotic chromosome must contain three elements: a centromere, telomeres, and numerous origins of replication.
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The DNA replication, transcription, and translation processes are intricately coupled in bacteria, allowing efficient gene expression and rapid protein synthesis. While this physical and functional coordination is advantageous, it introduces challenges that bacteria overcome through specific regulatory mechanisms.Coupling of Replication, Transcription, and TranslationThe coupling of replication, transcription, and translation is a hallmark of bacterial gene expression. As the replisome unwinds...
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Ribosomal DNA replication time coordinates completion of genome replication and anaphase in yeast.

Elizabeth X Kwan1, Gina M Alvino1, Kelsey L Lynch1

  • 1Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.

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Reducing the number of ribosomal DNA (rDNA) repeats in yeast causes rDNA to replicate early, delaying genome-wide replication. This triggers premature entry into anaphase by releasing the Cdc14 phosphatase.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Genome replication must complete on time for mitosis, genome integrity, and cell survival.
  • Organisms maintain hundreds of untranscribed ribosomal DNA (rDNA) copies, posing a challenge to timely replication.
  • rDNA arrays are replicated late in S phase due to their large size, repetitive nature, and essentiality.

Purpose of the Study:

  • To investigate the role of ribosomal DNA (rDNA) copy number in determining rDNA replication timing.
  • To explore the consequences of altered rDNA replication timing on genome-wide replication and cell cycle progression.
  • To elucidate the signaling mechanism linking rDNA replication completion to cell cycle progression.

Main Methods:

  • Utilized Saccharomyces cerevisiae (yeast) as a model organism.
  • Manipulated the number of rDNA repeats within the yeast genome.
  • Monitored rDNA replication timing and genome-wide replication profiles.
  • Assessed the release of the mitotic phosphatase Cdc14 from the nucleolus.
  • Observed entry into anaphase.

Main Results:

  • Reducing the number of rDNA repeats led to earlier replication of rDNA arrays.
  • Early rDNA replication caused a delay in replication of other genomic regions.
  • Cells with early-replicating rDNA and delayed genome-wide replication aberrantly released Cdc14 from the nucleolus.
  • These cells entered anaphase prematurely.

Conclusions:

  • rDNA copy number is a key determinant of rDNA replication timing.
  • The completion of rDNA replication acts as a signal for cell cycle progression, mediated by Cdc14 release.
  • Aberrant rDNA replication timing can disrupt genome-wide replication and lead to premature mitotic entry.