Novel approaches to study helicases using magnetic tweezers

Ruiz-Gutierrez, Nadia; Rieu, Martin; Ouellet, Jimmy; Allemand, Jean-François; Croquette, Vincent; Hir, Hervé Le

doi:10.1016/bs.mie.2022.03.035

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Mechanism Of Catalytic Activity1

Magnetic Tweezers1

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2022

2024

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Cited by 6 publications

(2 citation statements)

References 82 publications

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“…This is carried out by pulling single stranded DNA (ssDNA) around clusters of GNZs making conformational changes and generating two ssDNAs, as presented in Fig. 2 (b) [ 5 , 43 ]. However, to complete a helicase reaction GNPs should detach from ssDNA.…”

Section: Mechanism Of Catalytic Activitymentioning

confidence: 99%

Surface modification strategies and the functional mechanisms of gold nanozyme in biosensing and bioassay

Garehbaghi

Ashrafi

Adam

et al. 2023

Materials Today Bio

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Section: Mechanism Of Catalytic Activitymentioning

confidence: 99%

Surface modification strategies and the functional mechanisms of gold nanozyme in biosensing and bioassay

Garehbaghi

Ashrafi

Adam

et al. 2023

Materials Today Bio

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“…All our measurements were carried out using magnetic tweezers, which is a single-molecule force spectroscopy technique (24). They allow measuring extensions of single DNA hairpin molecules tethered to a surface on one end and on a magnetic bead on the other end, while a force is applied through a magnetic bead.…”

Section: Magnetic Tweezersmentioning

confidence: 99%

Strand switching mechanism of Pif1 helicase induced by its collision with a G-quadruplex embedded in dsDNA

Valle-Orero

Rieu

Tran

et al. 2022

Nucleic Acids Research

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G-rich sequences found at multiple sites throughout all genomes may form secondary structures called G-quadruplexes (G4), which act as roadblocks for molecular motors. Among the enzymes thought to process these structures, the Pif1 DNA helicase is considered as an archetypical G4-resolvase and its absence has been linked to G4-related genomic instabilities in yeast. Here we developed a single-molecule assay to observe Pif1 opening a DNA duplex and resolving the G4 in real time. In support of former enzymological studies, we show that the helicase reduces the lifetime of G4 from hours to seconds. However, we observe that in the presence of a G4, Pif1 exhibits a strong strand switching behavior, which can lead to Pif1 escaping G4 resolution, depending on the structural context surrounding the substrate. This behavior is also detected in the presence of other roadblocks (LNA or RNA). We propose that the efficiency of Pif1 to remove a roadblock (G4 or other) is affected by its strand switching behavior and depends on the context surrounding the obstacle. We discuss how this switching behavior may explain several aspects of Pif1 substrate preference and affect its activity as a G4 resolvase in vivo.

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Magnetic tweezers characterization of the entropic elasticity of intrinsically disordered proteins and peptoids

Truong,

Saleh

2024

Magnetic Tweezers for the Study of Protein Structure and Function

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Novel approaches to study helicases using magnetic tweezers

Cited by 6 publications

References 82 publications

Surface modification strategies and the functional mechanisms of gold nanozyme in biosensing and bioassay

Surface modification strategies and the functional mechanisms of gold nanozyme in biosensing and bioassay

Strand switching mechanism of Pif1 helicase induced by its collision with a G-quadruplex embedded in dsDNA

Magnetic tweezers characterization of the entropic elasticity of intrinsically disordered proteins and peptoids

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