Mapping DNA Topoisomerase Binding and Cleavage Genome Wide Using Next-Generation Sequencing Techniques
Next-generation sequencing (NGS) platforms have been adapted to generate genome-wide maps and sequence context of binding and cleavage of DNA topoisomerases (topos). Continuous refinements of these techniques have resulted in the acquisition of data with unprecedented depth and resolution, which has...
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doaj-43c25ac5da984537875befe7265169442020-11-25T01:45:17ZengMDPI AGGenes2073-44252020-01-011119210.3390/genes11010092genes11010092Mapping DNA Topoisomerase Binding and Cleavage Genome Wide Using Next-Generation Sequencing TechniquesShannon J. McKie0Anthony Maxwell1Keir C. Neuman2Laboratory of Single Molecule Biophysics, NHLBI, Bethesda, MD 20892, USADepartment of Biological Chemistry, John Innes Centre, Norwich NR4 7UH, UKLaboratory of Single Molecule Biophysics, NHLBI, Bethesda, MD 20892, USANext-generation sequencing (NGS) platforms have been adapted to generate genome-wide maps and sequence context of binding and cleavage of DNA topoisomerases (topos). Continuous refinements of these techniques have resulted in the acquisition of data with unprecedented depth and resolution, which has shed new light on in vivo topo behavior. Topos regulate DNA topology through the formation of reversible single- or double-stranded DNA breaks. Topo activity is critical for DNA metabolism in general, and in particular to support transcription and replication. However, the binding and activity of topos over the genome in vivo was difficult to study until the advent of NGS. Over and above traditional chromatin immunoprecipitation (ChIP)-seq approaches that probe protein binding, the unique formation of covalent protein−DNA linkages associated with DNA cleavage by topos affords the ability to probe cleavage and, by extension, activity over the genome. NGS platforms have facilitated genome-wide studies mapping the behavior of topos in vivo, how the behavior varies among species and how inhibitors affect cleavage. Many NGS approaches achieve nucleotide resolution of topo binding and cleavage sites, imparting an extent of information not previously attainable. We review the development of NGS approaches to probe topo interactions over the genome in vivo and highlight general conclusions and quandaries that have arisen from this rapidly advancing field of topoisomerase research.https://www.mdpi.com/2073-4425/11/1/92dna topoisomerasenext-generation sequencinggenome widetopoisomerase cleavagetopoisomerase bindingantibioticsanticancer drugs |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Shannon J. McKie Anthony Maxwell Keir C. Neuman |
spellingShingle |
Shannon J. McKie Anthony Maxwell Keir C. Neuman Mapping DNA Topoisomerase Binding and Cleavage Genome Wide Using Next-Generation Sequencing Techniques Genes dna topoisomerase next-generation sequencing genome wide topoisomerase cleavage topoisomerase binding antibiotics anticancer drugs |
author_facet |
Shannon J. McKie Anthony Maxwell Keir C. Neuman |
author_sort |
Shannon J. McKie |
title |
Mapping DNA Topoisomerase Binding and Cleavage Genome Wide Using Next-Generation Sequencing Techniques |
title_short |
Mapping DNA Topoisomerase Binding and Cleavage Genome Wide Using Next-Generation Sequencing Techniques |
title_full |
Mapping DNA Topoisomerase Binding and Cleavage Genome Wide Using Next-Generation Sequencing Techniques |
title_fullStr |
Mapping DNA Topoisomerase Binding and Cleavage Genome Wide Using Next-Generation Sequencing Techniques |
title_full_unstemmed |
Mapping DNA Topoisomerase Binding and Cleavage Genome Wide Using Next-Generation Sequencing Techniques |
title_sort |
mapping dna topoisomerase binding and cleavage genome wide using next-generation sequencing techniques |
publisher |
MDPI AG |
series |
Genes |
issn |
2073-4425 |
publishDate |
2020-01-01 |
description |
Next-generation sequencing (NGS) platforms have been adapted to generate genome-wide maps and sequence context of binding and cleavage of DNA topoisomerases (topos). Continuous refinements of these techniques have resulted in the acquisition of data with unprecedented depth and resolution, which has shed new light on in vivo topo behavior. Topos regulate DNA topology through the formation of reversible single- or double-stranded DNA breaks. Topo activity is critical for DNA metabolism in general, and in particular to support transcription and replication. However, the binding and activity of topos over the genome in vivo was difficult to study until the advent of NGS. Over and above traditional chromatin immunoprecipitation (ChIP)-seq approaches that probe protein binding, the unique formation of covalent protein−DNA linkages associated with DNA cleavage by topos affords the ability to probe cleavage and, by extension, activity over the genome. NGS platforms have facilitated genome-wide studies mapping the behavior of topos in vivo, how the behavior varies among species and how inhibitors affect cleavage. Many NGS approaches achieve nucleotide resolution of topo binding and cleavage sites, imparting an extent of information not previously attainable. We review the development of NGS approaches to probe topo interactions over the genome in vivo and highlight general conclusions and quandaries that have arisen from this rapidly advancing field of topoisomerase research. |
topic |
dna topoisomerase next-generation sequencing genome wide topoisomerase cleavage topoisomerase binding antibiotics anticancer drugs |
url |
https://www.mdpi.com/2073-4425/11/1/92 |
work_keys_str_mv |
AT shannonjmckie mappingdnatopoisomerasebindingandcleavagegenomewideusingnextgenerationsequencingtechniques AT anthonymaxwell mappingdnatopoisomerasebindingandcleavagegenomewideusingnextgenerationsequencingtechniques AT keircneuman mappingdnatopoisomerasebindingandcleavagegenomewideusingnextgenerationsequencingtechniques |
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