Novel approaches to study helicases using magnetic tweezers.

Autor:	Ruiz-Gutierrez N; Institut de Biologie de l'Ecole Normale Supérieure (IBENS), Ecole normale supérieure, CNRS, INSERM, PSL Research University, Paris, France., Rieu M; Institut de Biologie de l'Ecole Normale Supérieure (IBENS), Ecole normale supérieure, CNRS, INSERM, PSL Research University, Paris, France; Laboratoire de Physique de L'Ecole Normale Supérieure de Paris, CNRS, ENS, Université PSL, Sorbonne Université, Université de Paris, Paris, France., Ouellet J; Depixus SAS, 3-5 impasse Reille, Paris, France., Allemand JF; Institut de Biologie de l'Ecole Normale Supérieure (IBENS), Ecole normale supérieure, CNRS, INSERM, PSL Research University, Paris, France; Laboratoire de Physique de L'Ecole Normale Supérieure de Paris, CNRS, ENS, Université PSL, Sorbonne Université, Université de Paris, Paris, France., Croquette V; Institut de Biologie de l'Ecole Normale Supérieure (IBENS), Ecole normale supérieure, CNRS, INSERM, PSL Research University, Paris, France; Laboratoire de Physique de L'Ecole Normale Supérieure de Paris, CNRS, ENS, Université PSL, Sorbonne Université, Université de Paris, Paris, France; ESPCI Paris, Université PSL, Paris, France. Electronic address: vincent.croquette@espci.psl.eu., Le Hir H; Institut de Biologie de l'Ecole Normale Supérieure (IBENS), Ecole normale supérieure, CNRS, INSERM, PSL Research University, Paris, France. Electronic address: lehir@bio.ens.psl.eu.
Jazyk:	angličtina
Zdroj:	Methods in enzymology [Methods Enzymol] 2022; Vol. 673, pp. 359-403. Date of Electronic Publication: 2022 Apr 22.
DOI:	10.1016/bs.mie.2022.03.035
Abstrakt:	Helicases form a universal family of molecular motors that bind and translocate onto nucleic acids. They are involved in essentially every aspect of nucleic acid metabolism: from DNA replication to RNA decay, and thus ensure a large spectrum of functions in the cell, making their study essential. The development of micromanipulation techniques such as magnetic tweezers for the mechanistic study of these enzymes has provided new insights into their behavior and their regulation that were previously unrevealed by bulk assays. These experiments allowed very precise measures of their translocation speed, processivity and polarity. Here, we detail our newest technological advances in magnetic tweezers protocols for high-quality measurements and we describe the new procedures we developed to get a more profound understanding of helicase dynamics, such as their translocation in a force independent manner, their nucleic acid binding kinetics and their interaction with roadblocks. (Copyright © 2022 Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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