Comprehensive Identification of Cell Cycleregulated Genes of the Yeast Saccharomyces cerevisiae by M

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Comprehensive Identification of Cell
Cycle-regulated Genes of the Yeast Saccharomyces
cerevisiae by Microarray Hybridization

• Spellman et al.

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Questions

• What genes are cell cycle regulated?
• What genes are controlled by cell cycle
  regulators Cln3p and Clb2p?
• Are these cell cycle genes regulated by the same
  transcription factors?

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Cell Cycle
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Why is it important to study the cell cycle?

• Cancer
• Development
• Aging
• Microbes environment, disease
• (prokaryotes dont have a cell cycle like
  eukaryotes)

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Approach

• Genomics
• Three different synchronization methods
• a factor arrest
• cdc15 temperature-sensitive mutant
• Elutriation Induced expression of Cln3p and Clb2p
• Elutriation Induced expression of Cln3p and Clb2p
• DNA sequence analysis

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What genes are cell cycle regulated?

• Three independent synchronization methods.
• Took cell samples over two cell cycles.
• Data analysis was carried out in two different
  ways
• Phasing
• Clustering

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Phasing vs Clustering
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Major clusters
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G1 clusters

• CLN2 cluster genes involved in DNA replication
• Y cluster Y elements located in the telomeres
• Cluster of 92 genes involved with cell wall
  synthesis

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S and M clusters

• Histone Cluster
• MET cluster methionine biosynthesis
• CLB2 cluster genes involved in mitosis

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M/G1 Clusters

• MCM cluster genes involved in DNA replication
• SIC1 cluster genes that peak at the M/G1
  boundary
• MAT cluster genes that are MATa specific

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Induction of Cln3p and Clb2p

• 116 cell cycle regulated genes are induced by
  Cln3p and repressed by Clb2p
• 53 genes that are not cell cycle regulated
  respond to Cln3p or Clb2p
• Large overlap of genes regulated by these
  transcription factors

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DNA binding sites
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Conclusions

• 800 genes were found to be cell cycle regulated
• There are 500 genes that are regulated by know
  transcription factors
• That leaves 300 cell cycle genes whose
  regulation mechanism is unknown

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