Protein-induced Local DNA Bends Regulate Global Topology of Recombination Products

Quan Du, Alexei Livshits, Agnieszka Kwiatek, Makkuni Jayaram, Alexander Vologodskii

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)170-182
Number of pages13
JournalJournal of Molecular Biology
Volume368
Issue number1
DOIs
StatePublished - Apr 20 2007

Keywords

  • Cre
  • DNA topology
  • Flp
  • site-specific recombination
  • tyrosine family of recombinases

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Fingerprint Dive into the research topics of 'Protein-induced Local DNA Bends Regulate Global Topology of Recombination Products'. Together they form a unique fingerprint.

Cite this