Novel approaches to study helicases using magnetic tweezers.
Methods Enzymol
; 673: 359-403, 2022.
Article
en En
| MEDLINE
| ID: mdl-35965012
ABSTRACT
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.
Palabras clave
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Ácidos Nucleicos
/
ADN Helicasas
Idioma:
En
Revista:
Methods Enzymol
Año:
2022
Tipo del documento:
Article