Project description:Binding of transcription factors to DNA is mediated by the recognition of the chemical signatures of the DNA bases and the three-dimensional shape of the DNA molecule. The direct contribution of DNA shape to DNA-binding specificity has been difficult to assess, as DNA shape is a consequence of its sequence. Here, we teased apart these two modes of recognition in the context of Hox-DNA binding. We made a series of mutations in Hox residues that, in a co-crystal structure, only recognize DNA shape, and tested the effect on DNA binding preferences using SELEX-seq. Analysis of shape features of selected sequences revealed that these residues are both necessary and sufficient for selection of sequences with distinct shape features. We used statistical machine learning to show that the accuracy of binding specificity predictions improves by adding shape features to a model that only depends on sequence. We conclude that shape readout is a direct and critical component of binding site selection by Hox proteins. Three rounds of SELEX were performed on a series of Hox mutants as described in Slattery et al, Cell, 2011 (PMID 22153072) and Riley et al, Methods in molecular Biology, 2014 (PMID 25151169). Briefly, His-tagged Scr and Antp mutant proteins were incubated with a randomized 16mer oligonucleotide library, and bound DNA was amplified and sequenced as described (PMID 22153072, PMID 25151169).

2015-01-21 | E-GEOD-65073 | biostudies-arrayexpress

DNA binding factors shape the mouse methylome at distal regulatory regions.

Project description:This SuperSeries is composed of the following subset Series: GSE30202: DNA binding factors shape the mouse methylome at distal regulatory regions [BIS_seq] GSE30203: DNA binding factors shape the mouse methylome at distal regulatory regions [ChIP-seq] GSE30204: DNA binding factors shape the mouse methylome at distal regulatory regions [ChIP_chip] GSE30280: DNA binding factors shape the mouse methylome at distal regulatory regions [RNA_seq] Refer to individual Series

2011-12-09 | E-GEOD-30206 | biostudies-arrayexpress

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