High-intensity UV laser ChIP-seq for the study of protein-DNA interactions in living cells.

ABSTRACT: Genome-wide mapping of transcription factor binding is generally performed by chemical protein-DNA crosslinking, followed by chromatin immunoprecipitation and deep sequencing (ChIP-seq). Here we present the ChIP-seq technique based on photochemical crosslinking of protein-DNA interactions by high-intensity ultraviolet (UV) laser irradiation in living mammalian cells (UV-ChIP-seq). UV laser irradiation induces an efficient and instant formation of covalent "zero-length" crosslinks exclusively between nucleic acids and proteins that are in immediate contact, thus resulting in a "snapshot" of direct protein-DNA interactions in their natural environment. Here we show that UV-ChIP-seq, applied for genome-wide profiling of the sequence-specific transcriptional repressor B-cell lymphoma 6 (BCL6) in human diffuse large B-cell lymphoma (DLBCL) cells, produces sensitive and precise protein-DNA binding profiles, highly enriched with canonical BCL6 DNA sequence motifs. Using this technique, we also found numerous previously undetectable direct BCL6 binding sites, particularly in condensed, inaccessible areas of chromatin.

SUBMITTER: Steube A 

PROVIDER: S-EPMC5670203 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature