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Unravelling the mechanisms of Type 1A topoisomerases using single-molecule approaches

Topoisomerases are essential enzymes that regulate DNA topology. Type 1A family topoisomerases are found in nearly all living organisms and are unique in that they require single-stranded (ss)DNA for activity. These enzymes are vital for maintaining supercoiling homeostasis and resolving DNA entangl...

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Autores principales: Spakman, Dian, Bakx, Julia A M, Biebricher, Andreas S, Peterman, Erwin J G, Wuite, Gijs J L, King, Graeme A
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8191776/
https://www.ncbi.nlm.nih.gov/pubmed/33963870
http://dx.doi.org/10.1093/nar/gkab239
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author Spakman, Dian
Bakx, Julia A M
Biebricher, Andreas S
Peterman, Erwin J G
Wuite, Gijs J L
King, Graeme A
author_facet Spakman, Dian
Bakx, Julia A M
Biebricher, Andreas S
Peterman, Erwin J G
Wuite, Gijs J L
King, Graeme A
author_sort Spakman, Dian
collection PubMed
description Topoisomerases are essential enzymes that regulate DNA topology. Type 1A family topoisomerases are found in nearly all living organisms and are unique in that they require single-stranded (ss)DNA for activity. These enzymes are vital for maintaining supercoiling homeostasis and resolving DNA entanglements generated during DNA replication and repair. While the catalytic cycle of Type 1A topoisomerases has been long-known to involve an enzyme-bridged ssDNA gate that allows strand passage, a deeper mechanistic understanding of these enzymes has only recently begun to emerge. This knowledge has been greatly enhanced through the combination of biochemical studies and increasingly sophisticated single-molecule assays based on magnetic tweezers, optical tweezers, atomic force microscopy and Förster resonance energy transfer. In this review, we discuss how single-molecule assays have advanced our understanding of the gate opening dynamics and strand-passage mechanisms of Type 1A topoisomerases, as well as the interplay of Type 1A topoisomerases with partner proteins, such as RecQ-family helicases. We also highlight how these assays have shed new light on the likely functional roles of Type 1A topoisomerases in vivo and discuss recent developments in single-molecule technologies that could be applied to further enhance our understanding of these essential enzymes.
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spelling pubmed-81917762021-06-11 Unravelling the mechanisms of Type 1A topoisomerases using single-molecule approaches Spakman, Dian Bakx, Julia A M Biebricher, Andreas S Peterman, Erwin J G Wuite, Gijs J L King, Graeme A Nucleic Acids Res Survey and Summary Topoisomerases are essential enzymes that regulate DNA topology. Type 1A family topoisomerases are found in nearly all living organisms and are unique in that they require single-stranded (ss)DNA for activity. These enzymes are vital for maintaining supercoiling homeostasis and resolving DNA entanglements generated during DNA replication and repair. While the catalytic cycle of Type 1A topoisomerases has been long-known to involve an enzyme-bridged ssDNA gate that allows strand passage, a deeper mechanistic understanding of these enzymes has only recently begun to emerge. This knowledge has been greatly enhanced through the combination of biochemical studies and increasingly sophisticated single-molecule assays based on magnetic tweezers, optical tweezers, atomic force microscopy and Förster resonance energy transfer. In this review, we discuss how single-molecule assays have advanced our understanding of the gate opening dynamics and strand-passage mechanisms of Type 1A topoisomerases, as well as the interplay of Type 1A topoisomerases with partner proteins, such as RecQ-family helicases. We also highlight how these assays have shed new light on the likely functional roles of Type 1A topoisomerases in vivo and discuss recent developments in single-molecule technologies that could be applied to further enhance our understanding of these essential enzymes. Oxford University Press 2021-05-08 /pmc/articles/PMC8191776/ /pubmed/33963870 http://dx.doi.org/10.1093/nar/gkab239 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Survey and Summary
Spakman, Dian
Bakx, Julia A M
Biebricher, Andreas S
Peterman, Erwin J G
Wuite, Gijs J L
King, Graeme A
Unravelling the mechanisms of Type 1A topoisomerases using single-molecule approaches
title Unravelling the mechanisms of Type 1A topoisomerases using single-molecule approaches
title_full Unravelling the mechanisms of Type 1A topoisomerases using single-molecule approaches
title_fullStr Unravelling the mechanisms of Type 1A topoisomerases using single-molecule approaches
title_full_unstemmed Unravelling the mechanisms of Type 1A topoisomerases using single-molecule approaches
title_short Unravelling the mechanisms of Type 1A topoisomerases using single-molecule approaches
title_sort unravelling the mechanisms of type 1a topoisomerases using single-molecule approaches
topic Survey and Summary
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8191776/
https://www.ncbi.nlm.nih.gov/pubmed/33963870
http://dx.doi.org/10.1093/nar/gkab239
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