TY - JOUR
T1 - Protein-induced Local DNA Bends Regulate Global Topology of Recombination Products
AU - Du, Quan
AU - Livshits, Alexei
AU - Kwiatek, Agnieszka
AU - Jayaram, Makkuni
AU - Vologodskii, Alexander
N1 - Funding Information:
We thank the late Nick Cozzarelli for stimulating discussions on this work, and Paul Sadowski for providing Cre mutant plasmids and helpful advice. The work was supported by the National Institutes of Health grants GM54215 to A.V. and GM35654 to M.J. Partial support was provided by an award from the Robert F Welch Foundation to M.J. This investigation used a facility constructed with support from Research Facilities Improvement Grant Number C06 RR-16572-01 from the National Center for Research Resources, National Institutes of Health.
PY - 2007/4/20
Y1 - 2007/4/20
N2 - The tyrosine family of recombinases produces two smaller DNA circles when acting on circular DNA harboring two recombination sites in head-to-tail orientation. If the substrate is supercoiled, these circles can be unlinked or form multiply linked catenanes. The topological complexity of the products varies strongly even for similar recombination systems. This dependence has been solved here. Our computer simulation of the synapsis showed that the bend angles, φ{symbol}, created in isolated recombination sites by protein binding before assembly of the full complex, determine the product topology. To verify the validity of this theoretical finding we measured the values of φ{symbol} for Cre/loxP and Flp/FRT systems. The measurement was based on cyclization of the protein-bound short DNA fragments in solution. Despite the striking similarity of the synapses for these recombinases, action of Cre on head-to-tail target sites produces mainly unlinked circles, while that of Flp yields multiply linked catenanes. In full agreement with theoretical expectations we found that the values of φ{symbol} for these systems are very different, close to 35° and 80°, respectively. Our findings have general implications in how small protein machines acting locally on large DNA molecules exploit statistical properties of their substrates to bring about directed global changes in topology.
AB - The tyrosine family of recombinases produces two smaller DNA circles when acting on circular DNA harboring two recombination sites in head-to-tail orientation. If the substrate is supercoiled, these circles can be unlinked or form multiply linked catenanes. The topological complexity of the products varies strongly even for similar recombination systems. This dependence has been solved here. Our computer simulation of the synapsis showed that the bend angles, φ{symbol}, created in isolated recombination sites by protein binding before assembly of the full complex, determine the product topology. To verify the validity of this theoretical finding we measured the values of φ{symbol} for Cre/loxP and Flp/FRT systems. The measurement was based on cyclization of the protein-bound short DNA fragments in solution. Despite the striking similarity of the synapses for these recombinases, action of Cre on head-to-tail target sites produces mainly unlinked circles, while that of Flp yields multiply linked catenanes. In full agreement with theoretical expectations we found that the values of φ{symbol} for these systems are very different, close to 35° and 80°, respectively. Our findings have general implications in how small protein machines acting locally on large DNA molecules exploit statistical properties of their substrates to bring about directed global changes in topology.
KW - Cre
KW - DNA topology
KW - Flp
KW - site-specific recombination
KW - tyrosine family of recombinases
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U2 - 10.1016/j.jmb.2007.02.010
DO - 10.1016/j.jmb.2007.02.010
M3 - Article
C2 - 17337001
AN - SCOPUS:33947324808
SN - 0022-2836
VL - 368
SP - 170
EP - 182
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 1
ER -