5CNQ

Crystal structure of the Holliday junction-resolving enzyme GEN1 (WT) in complex with product DNA, Mg2+ and Mn2+ ions

Summary for 5CNQ

• Entry DOI: 10.2210/pdb5cnq/pdb
• Descriptor: Nuclease-like protein, R, DNA (5'-D(*TP*GP*AP*GP*CP*GP*GP*TP*GP*GP*TP*TP*GP*GP*T)-3'), ... (5 entities in total)
• Functional Keywords: gen1, 4-way holiday junction, resolvase, dna damage repair, replication
• Biological source: Chaetomium thermophilum; More
• Total number of polymer chains: 3
• Total formula weight: 61582.12

Authors

Liu, Y.J.,Freeman, A.D.J.,Declais, A.C.,Wilson, T.J.,Gartner, A.,Lilley, D.M.J. (deposition date: 2015-07-17, release date: 2015-12-30, Last modification date: 2024-10-16)

Primary citation

Liu, Y.,Freeman, A.D.,Declais, A.C.,Wilson, T.J.,Gartner, A.,Lilley, D.M.
Crystal Structure of a Eukaryotic GEN1 Resolving Enzyme Bound to DNA.
Cell Rep, 13:2565-2575, 2015

PubMed 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.

PubMed: 26686639
DOI: 10.1016/j.celrep.2015.11.042

Experimental method

X-RAY DIFFRACTION (2.602 Å)