Article abstract

Nature Methods 6, 283 - 289 (2009)
Published online: 22 March 2009 | doi:10.1038/nmeth.1313

Global mapping of protein-DNA interactions in vivo by digital genomic footprinting

Jay R Hesselberth1,6, Xiaoyu Chen2,6, Zhihong Zhang1,3,5, Peter J Sabo1, Richard Sandstrom1, Alex P Reynolds1, Robert E Thurman1, Shane Neph1, Michael S Kuehn1, William S Noble1,2, Stanley Fields1,3 & John A Stamatoyannopoulos1,4

The orchestrated binding of transcriptional activators and repressors to specific DNA sequences in the context of chromatin defines the regulatory program of eukaryotic genomes. We developed a digital approach to assay regulatory protein occupancy on genomic DNA in vivo by dense mapping of individual DNase I cleavages from intact nuclei using massively parallel DNA sequencing. Analysis of >23 million cleavages across the Saccharomyces cerevisiae genome revealed thousands of protected regulatory protein footprints, enabling de novo derivation of factor binding motifs and the identification of hundreds of new binding sites for major regulators. We observed striking correspondence between single-nucleotide resolution DNase I cleavage patterns and protein-DNA interactions determined by crystallography. The data also yielded a detailed view of larger chromatin features including positioned nucleosomes flanking factor binding regions. Digital genomic footprinting should be a powerful approach to delineate the cis-regulatory framework of any organism with an available genome sequence.

  1. Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
  2. Department of Computer Science, University of Washington, Seattle, Washington, USA.
  3. Howard Hughes Medical Institute, University of Washington, Seattle, Washington, USA.
  4. Department of Medicine, University of Washington, Seattle, Washington, USA.
  5. Present address: Illumina, Inc., San Diego, California, USA.
  6. These authors contributed equally to this work.

Correspondence to: John A Stamatoyannopoulos1,4 e-mail:


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