ArticleJournal of molecular biology2002
The order of strand exchanges in Cre-LoxP recombination and its basis suggested by the crystal structure of a Cre-LoxP Holliday junction complex.
Article in Journal of molecular biology, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 56 citations in OpenAlex.
- Engineering spacer specificity of the Cre/loxP system.Nucleic acids research · 2024Article
- Mechanisms of Cre recombinase synaptic complex assembly and activation illuminated by Cryo-EM.Nucleic acids research · 2022Article
- DNA binding induces aProceedings of the National Academy of Sciences of the United States of America · 2020Article
- Loop-closure kinetics reveal a stable, right-handed DNA intermediate in Cre recombination.Nucleic acids research · 2020Article
- Intricate and Cell Type-Specific Populations of Endogenous Circular DNA (eccDNA) inG3 (Bethesda, Md.) · 2017Article
- Xer Site Specific Recombination: Double and Single Recombinase Systems.Frontiers in microbiology · 2017Review
- Redesign of the monomer-monomer interface of Cre recombinase yields an obligate heterotetrameric complex.Nucleic acids research · 2015Article
- Insights into the preferential order of strand exchange in the Cre/loxP recombinase system: impact of the DNA spacer flanking sequence and flexibility.Journal of computer-aided molecular design · 2015Article
- Vika/vox, a novel efficient and specific Cre/loxP-like site-specific recombination system.Nucleic acids research · 2013Article
- Capturing reaction paths and intermediates in Cre-loxP recombination using single-molecule fluorescence.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- Homology-dependent interactions determine the order of strand exchange by IntDOT recombinase.Nucleic acids research · 2010Article
- A straight path to circular proteins.The Journal of biological chemistry · 2009Article
- Structure, dynamics, and branch migration of a DNA Holliday junction: a single-molecule fluorescence and modeling study.Biophysical journal · 2008Article
- Multiple levels of affinity-dependent DNA discrimination in Cre-LoxP recombination.Biochemistry · 2006Article
- Genome engineering in Bacillus anthracis using Cre recombinase.Infection and immunity · 2006Article
- Reversed DNA strand cleavage specificity in initiation of Cre-LoxP recombination induced by the His289Ala active-site substitution.Journal of molecular biology · 2005Article
- Preferential synapsis of loxP sites drives ordered strand exchange in Cre-loxP site-specific recombination.Nature chemical biology · 2005Article
- Mechanism of DNA compaction by yeast mitochondrial protein Abf2p.Biophysical journal · 2004Article
- A specificity switch in selected cre recombinase variants is mediated by macromolecular plasticity and water.Chemistry & biology · 2003Article
- Packaging of single DNA molecules by the yeast mitochondrial protein Abf2p.Biophysical journal · 2003Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
Cre recombinase uses two pairs of sequential cleavage and religation reactions to exchange homologous DNA strands between 34 base-pair (bp) LoxP recognition sequences. In the oligomeric recombination complex, a switch between "cleaving" and "non-cleaving" subunit conformations regulates the number, order, and regio-specificity of the strand exchanges. However, the particular sequence of events has been in question. From analysis of strand composition of the Holliday junction (HJ) intermediate, we determined that Cre initiates recombination of LoxP by cleaving the upper strand on the left arm. Cre preferred to react with the left arm of a LoxP suicide substrate, but at a similar rate to the right arm, indicating that the first strand to be exchanged is selected prior to cleavage. We propose that during complex assembly the cleaving subunit preferentially associates with the LoxP left arm, directing the first strand exchange to that side. In addition, this biased assembly would enforce productive orientation of LoxP sites in the recombination synapses. A novel Cre-HJ complex structure in which LoxP was oriented with the left arm bound by the cleaving Cre subunit suggested a physical basis for the strand exchange order. Lys86 and Lys201 interact with the left arm scissile adenine base differently than in structures that have a scissile guanine. These interactions are associated with positioning the 198-208 loop, a structural component of the conformational switch, in a configuration that is specific to the cleaving conformation. Our results suggest that strand exchange order and site alignment are regulated by an "induced fit" mechanism in which the cleaving conformation is selectively stabilized through protein-DNA interactions with the scissile base on the strand that is cleaved first.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.