Crystal Structure of a Eukaryotic GEN1 Resolving Enzyme Bound to DNA

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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 languageEnglish
Pages (from-to)2565-2575
Number of pages11
JournalCell Reports
Volume13
Issue number11
Early online date10 Dec 2015
DOIs
Publication statusPublished - 22 Dec 2015

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Crystal structure
DNA
Enzymes
2-Aminopurine
Crystal lattices
Dimers
Monomers
Fluorescence
Crystals
Proteins

Keywords

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

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",
author = "Yijin Liu and Freeman, {Alasdair D J} and D{\'e}clais, {Anne C{\'e}cile} and Wilson, {Timothy J.} and Anton Gartner and Lilley, {David M J}",
year = "2015",
month = "12",
day = "22",
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language = "English",
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TY - JOUR

T1 - Crystal Structure of a Eukaryotic GEN1 Resolving Enzyme Bound to DNA

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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U2 - 10.1016/j.celrep.2015.11.042

DO - 10.1016/j.celrep.2015.11.042

M3 - Article

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VL - 13

SP - 2565

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JO - Cell Reports

JF - Cell Reports

SN - 2211-1247

IS - 11

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