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Topo-Seq application for topoisomerases binding sites identification with a single-nucleotide resolution

ABSTRACT: Topo-Seq application for topoisomerases binding sites identification with a single-nucleotide resolution

PROVIDER: PRJNA481368 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR7527710_1.fastq.gz	Fastqsanger.gz
	SRR7527710_2.fastq.gz	Fastqsanger.gz
	SRR7527720_1.fastq.gz	Fastqsanger.gz
	SRR7527720_2.fastq.gz	Fastqsanger.gz
	SRR7527730_1.fastq.gz	Fastqsanger.gz

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Topo-Seq application for topoisomerases binding sites identification with a single-nucleotide resolution

Project description:Topo-Seq is a ChIP-Seq-based methodology that allows high-throughput identification of topoisomerases binding (cleavage) sites with a single-base precision. On a first stage of the project DNA-gyrase binding sites on a Escherichia coli DY330 genome are investigated.

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Project description:DNA topoisomerases are required to resolve DNA topological stress. Despite this essential role, abortive topoisomerase activity generates aberrant protein-linked DNA breaks, jeopardising genome stability. Here, to understand the genomic distribution and mechanisms underpinning topoisomerase-induced DNA breaks, we map Top2 DNA cleavage with strand-specific nucleotide resolution across the S. cerevisiae and human genomes—and use the meiotic Spo11 protein to validate the broad applicability of this method to explore the role of diverse topoisomerase family members. Our data characterises Mre11-dependent repair in yeast and defines two strikingly different fractions of Top2 activity in humans: tightly localised CTCF-proximal, and broadly distributed transcription-proximal, the latter correlated with gene length and expression. Moreover, single nucleotide accuracy reveals the influence primary DNA sequence has upon Top2 cleavage—distinguishing sites likely to form canonical DNA double-strand breaks (DSBs) from those predisposed to form strand-biased DNA single-strand breaks (SSBs) induced by etoposide (VP16) in vivo.

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