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Transcriptional regulatory code of a eukaryotic genome
Harbison CT et al. Nature, 2004, Vol. 431, pp. 99-104
Milica Volar3 March 2005.
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Outline of talk
• Background
• Yeast’s transcriptional regulatory map
• Promoter structure
• Regulator binding behaviour
• Summary
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Transcriptional regulators
• Proteins that bind to specific promoter sequences in genome
• Regulate genome expression and its maintenance
• Cell cycle, environment changes, replication, condensation, cohesion, etc.
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Procedure
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Genome wide location analysis
Crosslink protein to DNAin vivo with formaldehydeBreak open cells andshear DNA
Immunoprecipitate
Reverse-crosslinks,blunt DNA and ligateto unidirectional linkersLM-PCRLabeling
Hybridize to arrayRen B et al, 2004
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Procedure
They tested the binding by GWLA:
203 (84) DNA transcriptional regulators in rich media ->
11,000 unique interactions between regulators and promoter regions at high confidence (P<0.001)
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Yeast transcriptional regulatory map
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Yeast transcriptional regulatory map
3,353 interactions within 1,296 promoter regions
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Promoter architectures
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Promoter architectures
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Regulator binding behaviour
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Regulator binding behaviour
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Summary
•Construction of the yeast transcriptional regulatory map
•Promoter architectures
•Patterns of regulator binding behaviour