Search and processing of Holliday junctions within long DNA by junction-resolving enzymes.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
07 10 2022
07 10 2022
Historique:
received:
03
05
2022
accepted:
21
09
2022
entrez:
7
10
2022
pubmed:
8
10
2022
medline:
12
10
2022
Statut:
epublish
Résumé
Resolution of Holliday junctions is a critical intermediate step of homologous recombination in which junctions are processed by junction-resolving endonucleases. Although binding and cleavage are well understood, the question remains how the enzymes locate their substrate within long duplex DNA. Here we track fluorescent dimers of endonuclease I on DNA, presenting the complete single-molecule reaction trajectory for a junction-resolving enzyme finding and cleaving a Holliday junction. We show that the enzyme binds remotely to dsDNA and then undergoes 1D diffusion. Upon encountering a four-way junction, a catalytically-impaired mutant remains bound at that point. An active enzyme, however, cleaves the junction after a few seconds. Quantitative analysis provides a comprehensive description of the facilitated diffusion mechanism. We show that the eukaryotic junction-resolving enzyme GEN1 also undergoes facilitated diffusion on dsDNA until it becomes located at a junction, so that the general resolution trajectory is probably applicable to many junction resolving enzymes.
Identifiants
pubmed: 36207294
doi: 10.1038/s41467-022-33503-6
pii: 10.1038/s41467-022-33503-6
pmc: PMC9547003
doi:
Substances chimiques
DNA, Cruciform
0
DNA
9007-49-2
Endodeoxyribonucleases
EC 3.1.-
Endonucleases
EC 3.1.-
Holliday Junction Resolvases
EC 3.1.21.-
Deoxyribonuclease I
EC 3.1.21.1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
5921Subventions
Organisme : Medical Research Council
ID : FC0010048
Pays : United Kingdom
Organisme : Cancer Research UK
ID : A18604
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
Pays : United Kingdom
Organisme : Cancer Research UK
ID : 11722
Pays : United Kingdom
Organisme : Cancer Research UK
ID : FC0010048
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC-A658-5TY10
Pays : United Kingdom
Organisme : Wellcome Trust
ID : FC0010048
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 206292/Z/17/Z
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_UP_1102/5
Pays : United Kingdom
Informations de copyright
© 2022. The Author(s).
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