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

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

Molecules (Basel, Switzerland) (2021-08-28)
Roberto Fabian, Santosh Gaire, Christopher Tyson, Raghabendra Adhikari, Ian Pegg, Abhijit Sarkar
ABSTRACT

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.

MATERIALS
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Sigma-Aldrich
NAP1 Protein, active, 300 µg, Active N-terminal His6-tagged Drosophila NAP1 expressed in E. coli.