Sep 22, 2015

ChEC-seq kinetics discriminates transcription factor binding sites by DNA sequence and shape in vivo

BioRxiv : the Preprint Server for Biology
Gabriel E. ZentnerSteven Henikoff

Abstract

Chromatin endogenous cleavage (ChEC) uses fusion of a protein of interest to micrococcal nuclease (MNase) to target calcium-dependent cleavage to specific genomic loci in vivo. Here we report the combination of ChEC with high-throughput sequencing (ChEC-seq) to map budding yeast transcription factor (TF) binding. Temporal analysis of ChEC-seq data reveals two classes of sites for TFs, one displaying rapid cleavage at sites with robust consensus motifs and the second showing slow cleavage at largely unique sites with low-scoring motifs. Sites with high-scoring motifs also display asymmetric cleavage, indicating that ChEC-seq provides information on the directionality of TF-DNA interactions. Strikingly, similar DNA shape patterns are observed regardless of motif strength, indicating that the kinetics of ChEC-seq discriminates DNA recognition through sequence and/or shape. We propose that time-resolved ChEC-seq detects both high-affinity interactions of TFs with consensus motifs and sites preferentially sampled by TFs during diffusion and sliding.

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Mentioned in this Paper

In Vivo
Calcium [EPC]
Patterns
Micrococcal Nuclease
Genome
Calcium
Diffusion Weighted Imaging
Yeasts
Cytokinesis of the Fertilized Ovum
Sequence Determinations

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