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Cell Rep. 2015 Dec 22;13(11):2565-2575. doi: 10.1016/j.celrep.2015.11.042. Epub 2015 Dec 10.

Crystal Structure of a Eukaryotic GEN1 Resolving Enzyme Bound to DNA.

Author information

1
Cancer Research UK Nucleic Acid Structure Research Group, MSI/WTB Complex, University of Dundee, Dow Street, Dundee DD1 5EH, UK.
2
Center for Gene Regulation and Expression, MSI/WTB Complex, University of Dundee, Dow Street, Dundee DD1 5EH, UK.
3
Cancer Research UK Nucleic Acid Structure Research Group, MSI/WTB Complex, University of Dundee, Dow Street, Dundee DD1 5EH, UK. Electronic address: d.m.j.lilley@dundee.ac.uk.

Abstract

We present the crystal structure of the junction-resolving enzyme GEN1 bound to DNA at 2.5 Å resolution. The structure of the GEN1 protein reveals it to have an elaborated FEN-XPG family fold that is modified for its role in four-way junction resolution. The functional unit in the crystal is a monomer of active GEN1 bound to the product of resolution cleavage, with an extensive DNA binding interface for both helical arms. Within the crystal lattice, a GEN1 dimer interface juxtaposes two products, whereby they can be reconnected into a four-way junction, the structure of which agrees with that determined in solution. The reconnection requires some opening of the DNA structure at the center, in agreement with permanganate probing and 2-aminopurine fluorescence. The structure shows that a relaxation of the DNA structure accompanies cleavage, suggesting how second-strand cleavage is accelerated to ensure productive resolution of the junction.

KEYWORDS:

DNA recombination; DNA repair; DNA-protein interactions; Holliday junction; X-ray crystallography

PMID:
26686639
PMCID:
PMC4695337
DOI:
10.1016/j.celrep.2015.11.042
[Indexed for MEDLINE]
Free PMC Article

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