if i understand correctly, a dnase hotspot is a region/zone where dnaseI cuts, this is of variable lenght. Within a DNAse I hotspot there are DNAse I Sites (sometimes more than 1?) defined, or peaks,they are defined a fixed lenght (150bp?, FDR<0.01). I notice that a DGF peak is often the same as the HS peak, like here http://www.encodeproject.org/cgi-bin/hgTracks?hgsid=315808645 , i would expect the DGF peak to be inside the HS peak as a footprint is not sensitive to dnase I, could you comment on this?
I was exploring the encode content and am interested in DNAse I sites and prediction of transcription binding sites within dnase I sites. But there are 2 kinds of Dnase I tracks in UCS: a track called DNaseI Hypersensitivity by Digital DNaseI and a track Digital DNase Genomic Footprinting. What is the difference?
1 answer
The hypersensitivity track contains regions of DNaseI hypersensitivity (DHSs) called through a process developed in the Stamatoyannopolis lab. The Nature Methods paper by Sabo et al. describes this in more detail. The raw signal are windowed regions of chromatin accessibility across a genome, with the density of DNase cut fragments within each 150 bp window, every 20 bp. Peaks and hotspots are called from these regions.
Within nearly all DHSs, there is often one or more footprints found. This footprint is a relatively shorter region of variable length (6-40 bp, I think) where DNase does not cleave, because of bound proteins or protein complexes (or related phenomena, like lack of methylation, which allows proteins to bind DNA), like transcription factors or transcription initiation machinery. See the Nature ENCODE paper by Neph et al., also by the same lab, which explains this in more detail.
If you're doing TF prediction, the footprints will likely be of more use to you. Shane Neph's de novo motif discovery algorithm used these footprints to discover binding sites for 683 motifs. 394 of them matched entries in non-redundant and reduced known and experimental transcription factor databases (TRANSFAC. JASPAR Core, UniPROBE, and some data sets from the Kellis lab). The remaining 289 were found to be novel, frequented millions of footprints, and showed similarities with known TFs.
The differences are specifically: the DGF data are obtained from greater sequencing read depth than the HS set, and they contain footprint calls, whereas the HS data do not. There are also differences in the cell types published for both datasets, which I think was down to quality scores assigned to the sequencing results. The greater depth of coverage allows for picking up sensitive footprints which, as you note, are protected from DNaseI cleavage. If you want to know more about the hotspot calling algorithm, take a look here: http://www.uwencode.org/software/hotspot
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