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Related Experiment Videos

Chromosome XII context is important for rDNA function in yeast.

Yeon-Hee Kim1, Daisuke Ishikawa, Ho Phu Ha

  • 1Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.

Nucleic Acids Research
|June 2, 2006
PubMed
Summary

The chromosomal context of Saccharomyces cerevisiae chromosome XII is crucial for maintaining ribosomal DNA (rDNA) copy number and nucleolar function. Altering this context impacts rDNA stability, nucleolar structure, and organism lifespan.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • The ribosomal DNA (rDNA) locus in Saccharomyces cerevisiae is a large cluster of tandemly repeated units on chromosome XII.
  • The specific chromosomal context surrounding the rDNA cluster is hypothesized to influence its stability and function.
  • Understanding this context is key to comprehending rDNA homeostasis and its role in cellular physiology.

Purpose of the Study:

  • To investigate the biological significance of the chromosomal context of the rDNA cluster on chromosome XII.
  • To determine how deleting flanking regions of chromosome XII affects rDNA copy number, stability, and nucleolar morphology.
  • To assess the impact of altered chromosomal context on yeast lifespan and extrachromosomal rDNA circle formation.

Main Methods:

Related Experiment Videos

  • Engineered yeast strains with deleted variants of chromosome XII, isolating the rDNA cluster.
  • Quantified rDNA copy number in engineered strains.
  • Assessed the silencing frequency of URA3 inserted within the rDNA locus.
  • Examined nucleolar localization and morphology using Nop1p staining.
  • Measured yeast lifespan and quantified extrachromosomal rDNA circles.

Main Results:

  • A strain consisting solely of the rDNA cluster (1500 kb variant) showed a decreased rDNA copy number.
  • Increased URA3 gene silencing was observed in the rDNA-only strain.
  • Altered nucleolar morphology, including fewer Nop1p foci and a rounded shape, was noted.
  • Strains with deleted chromosome XII regions exhibited shorter lifespans and accumulated extrachromosomal rDNA circles.

Conclusions:

  • The chromosomal context of chromosome XII is essential for maintaining a stable rDNA copy number in Saccharomyces cerevisiae.
  • The specific genomic environment influences rDNA locus stability, gene silencing, and nucleolar organization.
  • Disruption of the native chromosomal context negatively impacts yeast physiology, leading to reduced lifespan and increased rDNA instability.