Scientific reports, 2016-04, Vol.6 (1), p.23697-23697, Article 23697
2016
Details
- Autor(en) / Beteiligte
- Titel
- Controlled rotation mechanism of DNA strand exchange by the Hin serine recombinase
- Ist Teil von
- Scientific reports, 2016-04, Vol.6 (1), p.23697-23697, Article 23697
- Ort / Verlag
- England: Nature Publishing Group
- Erscheinungsjahr
- 2016
- Link zum Volltext
- Quelle
- Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals
- Beschreibungen/Notizen
- DNA strand exchange by serine recombinases has been proposed to occur by a large-scale rotation of halves of the recombinase tetramer. Here we provide the first direct physical evidence for the subunit rotation mechanism for the Hin serine invertase. Single-DNA looping assays using an activated mutant (Hin-H107Y) reveal specific synapses between two hix sites. Two-DNA "braiding" experiments, where separate DNA molecules carrying a single hix are interwound, show that Hin-H107Y cleaves both hix sites and mediates multi-step rotational relaxation of the interwinding. The variable numbers of rotations in the DNA braid experiments are in accord with data from bulk experiments that follow DNA topological changes accompanying recombination by the hyperactive enzyme. The relatively slow Hin rotation rates, combined with pauses, indicate considerable rotary friction between synapsed subunit pairs. A rotational pausing mechanism intrinsic to serine recombinases is likely to be crucial for DNA ligation and for preventing deleterious DNA rearrangements.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 2045-2322eISSN: 2045-2322DOI: 10.1038/srep23697Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4817059
- Format
- –
- Schlagworte
- Bacterial Proteins - chemistry, Bacterial Proteins - genetics, Bacterial Proteins - metabolism, Deoxyribonucleic acid, DNA, DNA Nucleotidyltransferases - chemistry, DNA Nucleotidyltransferases - genetics, DNA Nucleotidyltransferases - metabolism, DNA, Superhelical - chemistry, DNA, Superhelical - metabolism, DNA-Binding Proteins - metabolism, Experiments, Factor For Inversion Stimulation Protein - metabolism, Models, Chemical, Models, Genetic, Models, Molecular, Nucleic Acid Conformation, Plasmids - genetics, Point Mutation, Protein Conformation, Protein Subunits, Recombinant Proteins - chemistry, Recombinant Proteins - metabolism, Recombinase, Recombination, Recombination, Genetic, Rotation, Salmonella - enzymology, Serine