1XO0

High resolution structure of the holliday junction intermediate in cre-loxp site-specific recombination

Summary for 1XO0

• Entry DOI: 10.2210/pdb1xo0/pdb
• Related: 1XNS 2CRX 3CRX
• Descriptor: loxP, Recombinase CRE, ... (4 entities in total)
• Functional Keywords: cre recombinase, holliday junction, recombination, complex (recombinase-dna), hydrolase, ligase-dna complex, ligase/dna
• Biological source: Enterobacteria phage P1
• Total number of polymer chains: 4
• Total formula weight: 94618.61

Authors

Ghosh, K.,Lau, C.K.,Guo, F.,Segall, A.M.,Van Duyne, G.D. (deposition date: 2004-10-05, release date: 2004-12-14, Last modification date: 2023-08-23)

Primary citation

Ghosh, K.,Lau, C.K.,Guo, F.,Segall, A.M.,Van Duyne, G.D.
Peptide trapping of the Holliday junction intermediate in Cre-loxP site-specific recombination.
J.Biol.Chem., 280:8290-8299, 2005

PubMed Abstract: Cre recombinase is a prototypical member of the tyrosine recombinase family of site-specific recombinases. Members of this family of enzymes catalyze recombination between specific DNA sequences by cleaving and exchanging one pair of strands between the two substrate sites to form a 4-way Holliday junction (HJ) intermediate and then resolve the HJ intermediate to recombinant products by a second round of strand exchanges. Recently, hexapeptide inhibitors have been described that are capable of blocking the second strand exchange step in the tyrosine recombinase recombination pathway, leading to an accumulation of the HJ intermediate. These peptides are active in the lambda-integrase, Cre recombinase, and Flp recombinase systems and are potentially important tools for both in vitro mechanistic studies and as in vivo probes of cellular function. Here we present biochemical and crystallographic data that support a model where the peptide inhibitor binds in the center of the recombinase-bound DNA junction and interacts with solvent-exposed bases near the junction branch point. Peptide binding induces large conformational changes in the DNA strands of the HJ intermediate, which affect the active site geometries in the recombinase subunits.

PubMed: 15591069
DOI: 10.1074/jbc.M411668200

Experimental method

X-RAY DIFFRACTION (2 Å)