Spatiotemporally controlled generation of NTPs for single-molecule studies

Sabantsev, Anton (author): Uppsala universitet,Molekylär systembiologi,Science for Life Laboratory, SciLifeLab

Mao, Guanzhong, 1985- (author): Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylär systembiologi

Aguirre, Javier Rivera, 1989- (author): Uppsala universitet,Molekylär systembiologi,Science for Life Laboratory, SciLifeLab

Panfilov, Mikhail (author): Peter Great St Petersburg Polytech Univ, St Petersburg, Russia.

Arseniev, Anatolii (author): Peter Great St Petersburg Polytech Univ, St Petersburg, Russia.;Russian Acad Sci, Ctr Precis Genome Editing & Genet Technol Biomed, Inst Gene Biol, Moscow, Russia.

Ho, Oanh (author): Uppsala Univ, Dept Cell & Mol Biol, Sci Life Lab, Uppsala, Sweden.

Khodorkovskiy, Mikhail (author): Peter Great St Petersburg Polytech Univ, St Petersburg, Russia.

Deindl, Sebastian (author): Uppsala universitet,Molekylär systembiologi,Science for Life Laboratory, SciLifeLab

(creator_code:org_t)

2022-09-21
2022
English.
In: Nature Chemical Biology. - : Springer Nature. - 1552-4450 .- 1552-4469. ; 18:10, s. 1144-

Related links:: https://doi.org/10.1...; show more...; https://uu.diva-port... (primary) (Raw object); https://urn.kb.se/re...; https://doi.org/10.1...; show less...

Abstract Subject headings

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.

By the author/editor: Sabantsev, Anton; Mao, Guanzhong, ...; Aguirre, Javier ...; Panfilov, Mikhai ...; Arseniev, Anatol ...; Ho, Oanh; show more...; Khodorkovskiy, M ...; Deindl, Sebastia ...; show less...

About the subject

NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...; and Biochemistry and ...

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