ArticleBiophysical journal2009
Tracking of single quantum dot labeled EcoRV sliding along DNA manipulated by double optical tweezers.
Article in Biophysical journal, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Single-trajectory Bayesian modeling reveals multi-state diffusion of the MSH sliding clamp.NPJ systems biology and applications · 2026Article
- Observing Protein One-Dimensional Sliding: Methodology and Biological Significance.Biomolecules · 2021Review
- Regulator-dependent temporal dynamics of a restriction-modification system's gene expression upon entering new host cells: single-cell and population studies.Nucleic acids research · 2021Article
- How p53 Molecules Solve the Target DNA Search Problem: A Review.International journal of molecular sciences · 2020Review
- Combining single-molecule manipulation and imaging for the study of protein-DNA interactions.Journal of visualized experiments : JoVE · 2014Article
- Type II restriction endonucleases--a historical perspective and more.Nucleic acids research · 2014Review
- Article
- Optical Methods to Study Protein-DNA Interactions in Vitro and in Living Cells at the Single-Molecule Level.International journal of molecular sciences · 2013Article
- DNA target sequence identification mechanism for dimer-active protein complexes.Nucleic acids research · 2013Article
- Future challenges in single-molecule fluorescence and laser trap approaches to studies of molecular motors.Developmental cell · 2012Review
- The rotation-coupled sliding of EcoRV.Nucleic acids research · 2012Article
- Dynamics of lesion processing by bacterial nucleotide excision repair proteins.Progress in molecular biology and translational science · 2012Review
- Labeling DNA for single-molecule experiments: methods of labeling internal specific sequences on double-stranded DNA.Nanoscale · 2011Review
- The diffusion constant of a labeled protein sliding along DNA.The European physical journal. E, Soft matter · 2011Article
- Efficient preparation of internally modified single-molecule constructs using nicking enzymes.Nucleic acids research · 2011Article
- Protein sliding and hopping kinetics on DNA.Physical review. E, Statistical, nonlinear, and soft matter physics · 2011Article
- Spatial epigenetic control of mono- and bistable gene expression.PLoS biology · 2010Article
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5 authors.
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Abstract
Fluorescence microscopy provides a powerful method to directly observe single enzymes moving along a DNA held in an extended conformation. In this work, we present results from single EcoRV enzymes labeled with quantum dots which interact with DNA manipulated by double optical tweezers. The application of quantum dots facilitated accurate enzyme tracking without photobleaching whereas the tweezers allowed us to precisely control the DNA extension. The labeling did not affect the biochemical activity of EcoRV checked by directly observing DNA digestion on the single molecule level. We used this system to demonstrate that during sliding, the enzyme stays in close contact with the DNA. Additionally, slight overstretching of the DNA resulted in a significant decrease of the 1D diffusion constant, which suggests that the deformation changes the energy landscape of the sliding interaction. Together with the simplicity of the setup, these results demonstrate that the combination of optical tweezers with fluorescence tracking is a powerful tool for the study of enzyme translocation along DNA.
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