Probing DNA - Transcription Factor Interactions Using Single-Molecule Fluorescence Detection in Nanofluidic Devices.
auxin response factor
lab-on-a-chip
microfluidics
single-molecule Förster resonance energy transfer
single-molecule biophysics
single-molecule fluorescence detection
total-internal reflection fluorescence microscopy
Journal
Advanced biology
ISSN: 2701-0198
Titre abrégé: Adv Biol (Weinh)
Pays: Germany
ID NLM: 101775319
Informations de publication
Date de publication:
04 2022
04 2022
Historique:
revised:
16
07
2021
received:
14
05
2021
pubmed:
3
9
2021
medline:
15
4
2022
entrez:
2
9
2021
Statut:
ppublish
Résumé
Single-molecule fluorescence detection offers powerful ways to study biomolecules and their complex interactions. Here, nanofluidic devices and camera-based, single-molecule Förster resonance energy transfer (smFRET) detection are combined to study the interactions between plant transcription factors of the auxin response factor (ARF) family and DNA oligonucleotides that contain target DNA response elements. In particular, it is shown that the binding of the unlabeled ARF DNA binding domain (ARF-DBD) to donor and acceptor labeled DNA oligonucleotides can be detected by changes in the FRET efficiency and changes in the diffusion coefficient of the DNA. In addition, this data on fluorescently labeled ARF-DBDs suggest that, at nanomolar concentrations, ARF-DBDs are exclusively present as monomers. In general, the fluidic framework of freely diffusing molecules minimizes potential surface-induced artifacts, enables high-throughput measurements, and proved to be instrumental in shedding more light on the interactions between ARF-DBDs monomers and between ARF-DBDs and their DNA response element.
Identifiants
pubmed: 34472724
doi: 10.1002/adbi.202100953
doi:
Substances chimiques
DNA Probes
0
Oligonucleotides
0
Transcription Factors
0
DNA
9007-49-2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2100953Informations de copyright
© 2021 The Authors. Advanced Biology published by Wiley-VCH GmbH.
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