Preparation and Resolution of Holliday Junction DNA Recombination Intermediates.
| Autor: | Shah Punatar R; The Francis Crick Institute, London, United Kingdom., West SC; The Francis Crick Institute, London, United Kingdom. Electronic address: stephen.west@crick.ac.uk. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in enzymology [Methods Enzymol] 2018; Vol. 600, pp. 569-590. Date of Electronic Publication: 2018 Jan 09. |
| DOI: | 10.1016/bs.mie.2017.11.022 |
| Abstrakt: | Holliday junctions provide a covalent link between recombining DNA molecules and need to be removed prior to chromosome segregation at mitosis. Defects in their resolution lead to mitotic catastrophe, characterized by the formation of DNA breaks and chromosome aberrations. Enzymes that resolve recombination intermediates have been identified in all forms of life, from bacteriophage, to bacteria, yeast, and humans. In higher eukaryotes, Holliday junctions are resolved by GEN1, a nuclease that is mechanistically similar to the prototypic resolvase Escherichia coli RuvC, and by the SMX trinuclease complex. Studies of these enzymes have been facilitated by the use of plasmid-sized DNA recombination intermediates made by RecA-mediated strand exchange. Here, we detail the preparation of these recombination intermediates, which resemble α-structures, and their resolution by RuvC and GEN1. (© 2018 Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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