期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 110, 期 43, 页码 17302-17307出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1311065110
关键词
tethered fluorophore motion; site-specific DNA recombination; protein-DNA interaction; chromosome segregation; single-molecule FRET
资金
- Wellcome Trust [WT083469MA]
- UK Engineering and Physical Sciences Research Council
- MathWorks
- European Commission [HEALTH-F4-2008-201418]
- Biotechnology and Biological Research Council [BB/H01795X/1]
- European Research Council [261227]
- European Research Council (ERC) [261227] Funding Source: European Research Council (ERC)
- Biotechnology and Biological Sciences Research Council [BB/H01795X/1] Funding Source: researchfish
- BBSRC [BB/H01795X/1] Funding Source: UKRI
Three single-molecule techniques have been used simultaneously and in tandem to track the formation in vitro of single XerCD-dif recombination complexes. We observed the arrival of the FtsK translocase at individual preformed synaptic complexes and demonstrated the conformational change that occurs during their activation. We then followed the reaction intermediate transitions as Holliday junctions formed through catalysis by XerD, isomerized, and were converted by XerC to reaction products, which then dissociated. These observations, along with the calculated intermediate lifetimes, inform the reaction mechanism, which plays a key role in chromosome unlinking in most bacteria with circular chromosomes.
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