transcriptional regulatory code of a eukaryotic genome harbison ct et al. nature, 2004, vol. 431,...

Transcriptional regulatory code of a eukaryotic genome

Harbison CT et al. Nature, 2004, Vol. 431, pp. 99-104

Milica Volar3 March 2005.

Outline of talk

• Background

• Yeast’s transcriptional regulatory map

• Promoter structure

• Regulator binding behaviour

• Summary

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.

Procedure

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

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)

Yeast transcriptional regulatory map

Yeast transcriptional regulatory map

3,353 interactions within 1,296 promoter regions

Promoter architectures

Promoter architectures

Regulator binding behaviour

Regulator binding behaviour

Summary

•Construction of the yeast transcriptional regulatory map

•Promoter architectures

•Patterns of regulator binding behaviour