PDBsum entry 5cnq

Replication

PDB id: 5cnq

Contents

Protein chain

392 a.a.

DNA/RNA

Metals

_MN ×2

Waters ×66

PDB id:

5cnq

Name:

Replication

Title:

Crystal structure of the holliday junction-resolving enzyme gen1 (wt) in complex with product DNA, mg2+ and mn2+ ions

Structure:

Nuclease-like protein. Chain: a. Fragment: catalytic domain, unp residues 2-465. Engineered: yes. R. Chain: r. Fragment: r-stem. Engineered: yes. DNA (5'-d( Tp Gp Ap Gp Cp Gp Gp Tp Gp Gp Tp Tp Gp Gp T)-

Source:

Chaetomium thermophilum. Organism_taxid: 209285. Gene: ctht_0007290. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Escherichia coli. Organism_taxid: 562. Other_details: DNA oligo synthesis.

Resolution:

2.60Å R-factor: 0.214 R-free: 0.252

Authors:

Y.J.Liu,A.D.J.Freeman,A.C.Declais,T.J.Wilson,A.Gartner,D.M.J.Lilley

Key ref:

Y.Liu et al. (2015). Crystal Structure of a Eukaryotic GEN1 Resolving Enzyme Bound to DNA. Cell Rep, 13, 2565-2575. PubMed id: 26686639 DOI: 10.1016/j.celrep.2015.11.042

Date:

17-Jul-15 Release date: 30-Dec-15

Protein chain

G0RYN2  (G0RYN2_CHATD) -  Nuclease-like protein from Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719)

Seq: 922 a.a.

Struc: 392 a.a.

DNA/RNA chains

T-A-C-C-C-A-C-C-A-C-C-G-C-T-C-A 16 bases

T-G-A-G-C-G-G-T-G-G-T-T-G-G-T 15 bases

DOI no: 10.1016/j.celrep.2015.11.042

Cell Rep 13:2565-2575 (2015)

PubMed id: 26686639

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

Y.Liu, A.D.Freeman, A.C.Déclais, T.J.Wilson, A.Gartner, D.M.Lilley.

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.