Journal
NATURE CHEMICAL BIOLOGY
Volume 18, Issue 10, Pages 1144-+Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41589-022-01100-9
Keywords
-
Categories
Funding
- European Research Council (ERC) Starting Grant [ERC-StG-714068]
- Knut and Alice Wallenberg Foundation [KAW/WAF 2019.0306, KAW 2016.0077]
- Cancerfonden [19 0055 Pj]
- Swedish Research Council [VR 03534]
- Ministry of Science and Higher Education of the Russian Federation [075-15-2021-1333]
Ask authors/readers for more resources
The study introduces a method called LAGOON for controlling NTP-driven reactions in single-molecule experiments by locally generating NTPs. This method significantly increases measurement throughput and enables single-turnover observations. The effectiveness of LAGOON is demonstrated in single-molecule fluorescence and force spectroscopy assays by monitoring DNA unwinding, nucleosome sliding, and RNA polymerase elongation. LAGOON can be easily integrated with various single-molecule techniques, and is expected to facilitate studies on a wide range of crucial NTP-driven processes.
Many essential processes in the cell depend on proteins that use nucleoside triphosphates (NTPs). Methods that directly monitor the often-complex dynamics of these proteins at the single-molecule level have helped to uncover their mechanisms of action. However, the measurement throughput is typically limited for NTP-utilizing reactions, and the quantitative dissection of complex dynamics over multiple sequential turnovers remains challenging. Here we present a method for controlling NTP-driven reactions in single-molecule experiments via the local generation of NTPs (LAGOON) that markedly increases the measurement throughput and enables single-turnover observations. We demonstrate the effectiveness of LAGOON in single-molecule fluorescence and force spectroscopy assays by monitoring DNA unwinding, nucleosome sliding and RNA polymerase elongation. LAGOON can be readily integrated with many single-molecule techniques, and we anticipate that it will facilitate studies of a wide range of crucial NTP-driven processes.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available