From Ensemble FRET to Single-Molecule Imaging: Monitoring Individual Cellular Machinery in Action

Islam, Farhana; Basu, Manali; Mishra, Padmaja P.

doi:10.1007/978-981-16-4550-1_6

Cited by 1 publication

(1 citation statement)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Fluorescence resonance energy transfer (FRET) is the phenomenon that involves the nonradiative energy transfer from a donor (D) fluorophore at its excited state to an acceptor (A, another fluorophore or a quencher) in the ground state due to dipole–dipole coupling when the appropriate spectral overlap is satisfied between the two fluorophores (donor and acceptor) in close proximity. Upon transferring a part of its energy to the acceptor, the donor’s fluorescence emission intensity falls with a subsequent rise in the acceptor’s fluorescence intensity.…”

Section: Single-molecule Fluorescence Resonance Energy Transfer (Smfret)mentioning

confidence: 99%

Insights into the Dynamics and Helicase Activity of RecD2 of Deinococcus radiodurans during DNA Repair: A Single-Molecule Perspective

2023

Self Cite

View full text Add to dashboard Cite

While the double helix is the most stable conformation of DNA inside cells, its transient unwinding and subsequent partial separation of the two complementary strands yields an intermediate single-stranded DNA (ssDNA). The ssDNA is involved in all major DNA transactions such as replication, transcription, recombination, and repair. The process of DNA unwinding and translocation is shouldered by helicases that transduce the chemical energy derived from nucleotide triphosphate (NTP) hydrolysis to mechanical energy and utilize it to destabilize hydrogen bonds between complementary base pairs. Consequently, a comprehensive understanding of the molecular mechanisms of these enzymes is essential. In the last few decades, a combination of single-molecule techniques (force-based manipulation and visualization) have been employed to study helicase action. These approaches have allowed researchers to study the single helicase−DNA complex in real-time and the free energy landscape of their interaction together with the detection of conformational intermediates and molecular heterogeneity during the course of helicase action. Furthermore, the unique ability of these techniques to resolve helicase motion at nanometer and millisecond spatial and temporal resolutions, respectively, provided surprising insights into their mechanism of action. This perspective outlines the contribution of single-molecule methods in deciphering molecular details of helicase activities. It also exemplifies how each technique was or can be used to study the helicase action of RecD2 in recombination DNA repair.

show abstract

Section: Single-molecule Fluorescence Resonance Energy Transfer (Smfret)mentioning

confidence: 99%

Insights into the Dynamics and Helicase Activity of RecD2 of Deinococcus radiodurans during DNA Repair: A Single-Molecule Perspective

2023

Self Cite

View full text Add to dashboard Cite

show abstract

From Ensemble FRET to Single-Molecule Imaging: Monitoring Individual Cellular Machinery in Action

Cited by 1 publication

References 27 publications

Insights into the Dynamics and Helicase Activity of RecD2 of Deinococcus radiodurans during DNA Repair: A Single-Molecule Perspective

Insights into the Dynamics and Helicase Activity of RecD2 of Deinococcus radiodurans during DNA Repair: A Single-Molecule Perspective

Contact Info

Product

Resources

About