Probing the mechanical stability of bridged DNA-H-NS protein complexes by single-molecule AFM pulling

Yan Liang, Ramon A. van der Valk, Remus T. Dame, Wouter H. Roos, Gijs J. L. Wuite

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Atomic force microscopy (AFM) has proven to be a powerful tool for the study of DNA-protein interactions due to its ability to image single molecules at the nanoscale. However, the use of AFM in force spectroscopy to study DNA-protein interactions has been limited. Here we developed a high throughput, AFM based, pulling assay to measure the strength and kinetics of protein bridging of DNA molecules. As a model system, we investigated the interactions between DNA and the Histone-like Nucleoid-Structuring protein (H-NS). We confirmed that H-NS both changes DNA rigidity and forms bridges between DNA molecules. This straightforward methodology provides a high-throughput approach with single-molecule resolution which is widely applicable to study cross-substrate interactions such as DNA-bridging proteins.
Original languageEnglish
Article number15275
JournalScientific Reports
Volume7
DOIs
Publication statusPublished - 10 Nov 2017

Cite this

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title = "Probing the mechanical stability of bridged DNA-H-NS protein complexes by single-molecule AFM pulling",
abstract = "Atomic force microscopy (AFM) has proven to be a powerful tool for the study of DNA-protein interactions due to its ability to image single molecules at the nanoscale. However, the use of AFM in force spectroscopy to study DNA-protein interactions has been limited. Here we developed a high throughput, AFM based, pulling assay to measure the strength and kinetics of protein bridging of DNA molecules. As a model system, we investigated the interactions between DNA and the Histone-like Nucleoid-Structuring protein (H-NS). We confirmed that H-NS both changes DNA rigidity and forms bridges between DNA molecules. This straightforward methodology provides a high-throughput approach with single-molecule resolution which is widely applicable to study cross-substrate interactions such as DNA-bridging proteins.",
author = "Yan Liang and {van der Valk}, {Ramon A.} and Dame, {Remus T.} and Roos, {Wouter H.} and Wuite, {Gijs J. L.}",
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doi = "10.1038/s41598-017-15477-4",
language = "English",
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Probing the mechanical stability of bridged DNA-H-NS protein complexes by single-molecule AFM pulling. / Liang, Yan; van der Valk, Ramon A.; Dame, Remus T.; Roos, Wouter H.; Wuite, Gijs J. L.

In: Scientific Reports, Vol. 7, 15275, 10.11.2017.

Research output: Contribution to JournalArticleAcademicpeer-review

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T1 - Probing the mechanical stability of bridged DNA-H-NS protein complexes by single-molecule AFM pulling

AU - Liang, Yan

AU - van der Valk, Ramon A.

AU - Dame, Remus T.

AU - Roos, Wouter H.

AU - Wuite, Gijs J. L.

PY - 2017/11/10

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AB - Atomic force microscopy (AFM) has proven to be a powerful tool for the study of DNA-protein interactions due to its ability to image single molecules at the nanoscale. However, the use of AFM in force spectroscopy to study DNA-protein interactions has been limited. Here we developed a high throughput, AFM based, pulling assay to measure the strength and kinetics of protein bridging of DNA molecules. As a model system, we investigated the interactions between DNA and the Histone-like Nucleoid-Structuring protein (H-NS). We confirmed that H-NS both changes DNA rigidity and forms bridges between DNA molecules. This straightforward methodology provides a high-throughput approach with single-molecule resolution which is widely applicable to study cross-substrate interactions such as DNA-bridging proteins.

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