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A Horizontal Magnetic Tweezers for Studying Single DNA Molecules and DNA-Binding Proteins

We report data from single molecule studies on the interaction between single DNA molecules and core histones using custom-designed horizontal magnetic tweezers. The DNA-core histone complexes were formed using λ-DNA tethers, core histones, and NAP1 and were exposed to forces ranging from ~2 pN to ~...

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Autores principales: Fabian, Roberto, Gaire, Santosh, Tyson, Christopher, Adhikari, Raghabendra, Pegg, Ian, Sarkar, Abhijit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8398817/
https://www.ncbi.nlm.nih.gov/pubmed/34443369
http://dx.doi.org/10.3390/molecules26164781
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author Fabian, Roberto
Gaire, Santosh
Tyson, Christopher
Adhikari, Raghabendra
Pegg, Ian
Sarkar, Abhijit
author_facet Fabian, Roberto
Gaire, Santosh
Tyson, Christopher
Adhikari, Raghabendra
Pegg, Ian
Sarkar, Abhijit
author_sort Fabian, Roberto
collection PubMed
description We report data from single molecule studies on the interaction between single DNA molecules and core histones using custom-designed horizontal magnetic tweezers. The DNA-core histone complexes were formed using λ-DNA tethers, core histones, and NAP1 and were exposed to forces ranging from ~2 pN to ~74 pN. During the assembly events, we observed the length of the DNA decrease in approximate integer multiples of ~50 nm, suggesting the binding of the histone octamers to the DNA tether. During the mechanically induced disassembly events, we observed disruption lengths in approximate integer multiples of ~50 nm, suggesting the unbinding of one or more octamers from the DNA tether. We also observed histone octamer unbinding events at forces as low as ~2 pN. Our horizontal magnetic tweezers yielded high-resolution, low-noise data on force-mediated DNA-core histone assembly and disassembly processes.
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spelling pubmed-83988172021-08-29 A Horizontal Magnetic Tweezers for Studying Single DNA Molecules and DNA-Binding Proteins Fabian, Roberto Gaire, Santosh Tyson, Christopher Adhikari, Raghabendra Pegg, Ian Sarkar, Abhijit Molecules Article We report data from single molecule studies on the interaction between single DNA molecules and core histones using custom-designed horizontal magnetic tweezers. The DNA-core histone complexes were formed using λ-DNA tethers, core histones, and NAP1 and were exposed to forces ranging from ~2 pN to ~74 pN. During the assembly events, we observed the length of the DNA decrease in approximate integer multiples of ~50 nm, suggesting the binding of the histone octamers to the DNA tether. During the mechanically induced disassembly events, we observed disruption lengths in approximate integer multiples of ~50 nm, suggesting the unbinding of one or more octamers from the DNA tether. We also observed histone octamer unbinding events at forces as low as ~2 pN. Our horizontal magnetic tweezers yielded high-resolution, low-noise data on force-mediated DNA-core histone assembly and disassembly processes. MDPI 2021-08-07 /pmc/articles/PMC8398817/ /pubmed/34443369 http://dx.doi.org/10.3390/molecules26164781 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Fabian, Roberto
Gaire, Santosh
Tyson, Christopher
Adhikari, Raghabendra
Pegg, Ian
Sarkar, Abhijit
A Horizontal Magnetic Tweezers for Studying Single DNA Molecules and DNA-Binding Proteins
title A Horizontal Magnetic Tweezers for Studying Single DNA Molecules and DNA-Binding Proteins
title_full A Horizontal Magnetic Tweezers for Studying Single DNA Molecules and DNA-Binding Proteins
title_fullStr A Horizontal Magnetic Tweezers for Studying Single DNA Molecules and DNA-Binding Proteins
title_full_unstemmed A Horizontal Magnetic Tweezers for Studying Single DNA Molecules and DNA-Binding Proteins
title_short A Horizontal Magnetic Tweezers for Studying Single DNA Molecules and DNA-Binding Proteins
title_sort horizontal magnetic tweezers for studying single dna molecules and dna-binding proteins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8398817/
https://www.ncbi.nlm.nih.gov/pubmed/34443369
http://dx.doi.org/10.3390/molecules26164781
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