Imperial College London
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Professor David S. Rueda

Faculty of Medicine, Department of Infectious Disease

Chair in Molecular and Cellular Biophysics
 
 
 
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Contact

 

david.rueda Website

 
 
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Location

 

6.12DLMS BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Kaczmarczyk:2022:10.1038/s41467-022-33503-6,
author = {Kaczmarczyk, AP and Déclais, A-C and Newton, MD and Boulton, SJ and Lilley, DMJ and Rueda, DS},
doi = {10.1038/s41467-022-33503-6},
journal = {Nature Communications},
pages = {1--13},
title = {Search and processing of Holliday junctions within long DNA by junction-resolving enzymes},
url = {http://dx.doi.org/10.1038/s41467-022-33503-6},
volume = {13},
year = {2022}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - 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.
AU  - Kaczmarczyk,AP
AU  - Déclais,A-C
AU  - Newton,MD
AU  - Boulton,SJ
AU  - Lilley,DMJ
AU  - Rueda,DS
DO  - 10.1038/s41467-022-33503-6
EP  - 13
PY  - 2022///
SP  - 1
TI  - Search and processing of Holliday junctions within long DNA by junction-resolving enzymes
T2  - Nature Communications
UR  - http://dx.doi.org/10.1038/s41467-022-33503-6
UR  - https://www.nature.com/articles/s41467-022-33503-6
UR  - http://hdl.handle.net/10044/1/100020
VL  - 13
ER  -
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