Crystal Structure of a Eukaryotic GEN1 Resolving Enzyme Bound to DNA

Yijin Liu; Alasdair D J Freeman; Anne Cécile Déclais; Timothy J. Wilson; Anton Gartner; David M J Lilley

doi:10.1016/j.celrep.2015.11.042

Crystal Structure of a Eukaryotic GEN1 Resolving Enzyme Bound to DNA

Yijin Liu, Alasdair D J Freeman, Anne Cécile Déclais, Timothy J. Wilson, Anton Gartner, David M J Lilley (Lead / Corresponding author)

Research output: Contribution to journal › Article › peer-review

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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.

Original language	English
Pages (from-to)	2565-2575
Number of pages	11
Journal	Cell Reports
Volume	13
Issue number	11
Early online date	10 Dec 2015
DOIs	https://doi.org/10.1016/j.celrep.2015.11.042
Publication status	Published - 22 Dec 2015

Keywords

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

ASJC Scopus subject areas

General Biochemistry,Genetics and Molecular Biology

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10.1016/j.celrep.2015.11.042Licence: CC BY-NC-ND

Publisher final version
© 2015 The Authors. Published by Elsevier Inc. Open Access funded by Cancer Research UK. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)
Final published version, 3.89 MBLicence: CC BY-NC-ND

Cite this

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title = "Crystal Structure of a Eukaryotic GEN1 Resolving Enzyme Bound to DNA",

abstract = "We present the crystal structure of the junction-resolving enzyme GEN1 bound to DNA at 2.5 {\AA} 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",

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AU - Liu, Yijin

AU - Freeman, Alasdair D J

AU - Déclais, Anne Cécile

AU - Wilson, Timothy J.

AU - Gartner, Anton

AU - Lilley, David M J

PY - 2015/12/22

Y1 - 2015/12/22

N2 - 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.

AB - 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.

KW - DNA recombination

KW - DNA repair

KW - DNA-protein interactions

KW - Holliday junction

KW - X-ray crystallography

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Crystal Structure of a Eukaryotic GEN1 Resolving Enzyme Bound to DNA

Abstract

Keywords

ASJC Scopus subject areas

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Combined Genetic and Biochemical Approaches to Uncover and Characterize Redundant Factors Involved in Late Stages of Recombinational Repair

Lilley, David

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Crystal Structure of a Eukaryotic GEN1 Resolving Enzyme Bound to DNA

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

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Combined Genetic and Biochemical Approaches to Uncover and Characterize Redundant Factors Involved in Late Stages of Recombinational Repair

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Lilley, David

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