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- W1994448441 abstract "Site-specific recombination promotes integration, excision or inversion of DNA strands during gene transcription and chromosome replication. The recombination process generally involves the formation of synaptic complexes, DNA cleavage and rejoining, though the specific mechanism of each enzyme varies. The Hin recombinase is a member of the serine recombinase family which has an active serine residue that can catalyze the DNA cleavage and re-ligating. Hin functions to invert a 900 bp DNA segment between two hin-binding (hix) sites within the Salmonella chromosome that contains a promoter for downstream flagellar genes. The inversion requires the auxiliary protein Fis and two Fis-binding sites on the targeted DNA segment, which assemble together with hix sites and Hin to form an invertasome structure. The assembly is stimulated by the protein HU. A class of engineered Hin mutants Hin-H107Y (H107Y) is able to assemble oligonucleotide substrates containing hix sites into stable synaptic complexes that catalyze recombination without Fis and HU. Structural data suggest that Hin share with gamma-delta resolvase a similar helix-turn-helix domain at the 40-50 amino acid residues. We hypothesized that H107Y could act like resolvase to form four subunits synapse, cleave both strands of DNA within the center of hix site, rotate and rejoin. We therefore used single DNA assays to analyze the dynamics associated with H107Y catalyzed strand exchange. Enzyme-DNA binding and synapse formation was observed with a single DNA containing two hix sites. We further obtained data for subunits rotation and re-ligation from a braiding assay which consisted of double DNAs each having a single hix site, indicating that the rotation strand exchange mechanism observed recently for Bxb1 Int is also possessed by Hin." @default.
- W1994448441 created "2016-06-24" @default.
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- W1994448441 date "2013-01-01" @default.
- W1994448441 modified "2023-10-16" @default.
- W1994448441 title "Observation of Synapse Rotation of Engineered Recombinase HIN-H107Y" @default.
- W1994448441 doi "https://doi.org/10.1016/j.bpj.2012.11.2047" @default.
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