Journal article
DNA Topology and Geometry in Flp and Cre Recombination
Journal of molecular biology, Vol.357(4), pp.1089-1104
2006
DOI: 10.1016/j.jmb.2006.01.037
PMID: 16483600
Abstract
The Flp recombinase of yeast and the Cre recombinase of bacteriophage P1 both belong to the λ-integrase (Int) family of site-specific recombinases. These recombination systems recognize recombination-target sequences that consist of two 13
bp inverted repeats flanking a 6 or 8
bp spacer sequence. Recombination reactions involve particular geometric and topological relationships between DNA target sites at synapsis, which we investigate using nicked-circular DNA molecules. Examination of the tertiary structure of synaptic complexes formed on nicked plasmid DNAs by atomic-force microscopy, in conjunction with detailed topological analysis using the mathematics of tangles, shows that only a limited number of recombination-site topologies are consistent with the global structures of plasmids bearing directly and inversely repeated sites. The tangle solutions imply that there is significant distortion of the Holliday-junction intermediate relative to the planar structure of the four-way DNA junction present in the Flp and Cre co-crystal structures. Based on simulations of nucleoprotein structures that connect the two-dimensional tangle solutions with three-dimensional models of the complexes, we propose a recombination mechanism in which the synaptic intermediate is characterized by a non-planar, possibly near-tetrahedral, Holliday-junction intermediate. Only modest conformational changes within this structure are needed to form the symmetric, planar DNA junction, which may be characteristic of shorter-lived intermediates along the recombination pathway.
Details
- Title: Subtitle
- DNA Topology and Geometry in Flp and Cre Recombination
- Creators
- Alexandre A Vetcher - The University of Texas at DallasAlexander Y Lushnikov - University of Nebraska Medical CenterJunalyn Navarra-Madsen - Texas Woman's UniversityRobert G SchareinYuri L Lyubchenko - University of Nebraska Medical CenterIsabel K Darcy - University of IowaStephen D Levene - The University of Texas at Dallas
- Resource Type
- Journal article
- Publication Details
- Journal of molecular biology, Vol.357(4), pp.1089-1104
- DOI
- 10.1016/j.jmb.2006.01.037
- PMID
- 16483600
- NLM abbreviation
- J Mol Biol
- ISSN
- 0022-2836
- eISSN
- 1089-8638
- Publisher
- Elsevier Ltd
- Language
- English
- Date published
- 2006
- Academic Unit
- Mathematics
- Record Identifier
- 9984241148602771
Metrics
16 Record Views