Sequence-specific dynamic DNA bending explains mitochondrial TFAM's dual role in DNA packaging and transcription initiation.
DNA-Binding Proteins
/ metabolism
Mitochondrial Proteins
/ metabolism
Transcription Factors
/ metabolism
DNA, Mitochondrial
/ genetics
DNA Packaging
Promoter Regions, Genetic
Fluorescence Resonance Energy Transfer
Humans
Nucleic Acid Conformation
Transcription Initiation, Genetic
Mitochondria
/ metabolism
Single Molecule Imaging
DNA-Directed RNA Polymerases
/ metabolism
Base Sequence
Protein Binding
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
27 Jun 2024
27 Jun 2024
Historique:
received:
06
09
2023
accepted:
17
06
2024
medline:
28
6
2024
pubmed:
28
6
2024
entrez:
27
6
2024
Statut:
epublish
Résumé
Mitochondrial transcription factor A (TFAM) employs DNA bending to package mitochondrial DNA (mtDNA) into nucleoids and recruit mitochondrial RNA polymerase (POLRMT) at specific promoter sites, light strand promoter (LSP) and heavy strand promoter (HSP). Herein, we characterize the conformational dynamics of TFAM on promoter and non-promoter sequences using single-molecule fluorescence resonance energy transfer (smFRET) and single-molecule protein-induced fluorescence enhancement (smPIFE) methods. The DNA-TFAM complexes dynamically transition between partially and fully bent DNA conformational states. The bending/unbending transition rates and bending stability are DNA sequence-dependent-LSP forms the most stable fully bent complex and the non-specific sequence the least, which correlates with the lifetimes and affinities of TFAM with these DNA sequences. By quantifying the dynamic nature of the DNA-TFAM complexes, our study provides insights into how TFAM acts as a multifunctional protein through the DNA bending states to achieve sequence specificity and fidelity in mitochondrial transcription while performing mtDNA packaging.
Identifiants
pubmed: 38937458
doi: 10.1038/s41467-024-49728-6
pii: 10.1038/s41467-024-49728-6
doi:
Substances chimiques
DNA-Binding Proteins
0
Mitochondrial Proteins
0
Transcription Factors
0
DNA, Mitochondrial
0
TFAM protein, human
0
DNA-Directed RNA Polymerases
EC 2.7.7.6
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
5446Subventions
Organisme : U.S. Department of Energy (DOE)
ID : DE-SC0019313
Organisme : U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (NIGMS)
ID : GM118086
Informations de copyright
© 2024. The Author(s).
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