Assembly, translocation, and activation of XerCD-dif recombination by FtsK translocase analyzed in real-time by FRET and two-color tethered fluorophore motion

Proceedings of the National Academy of Sciences of the United States of America
Peter F J MayLidia K Arciszewska

Abstract

The FtsK dsDNA translocase functions in bacterial chromosome unlinking by activating XerCD-dif recombination in the replication terminus region. To analyze FtsK assembly and translocation, and the subsequent activation of XerCD-dif recombination, we extended the tethered fluorophore motion technique, using two spectrally distinct fluorophores to monitor two effective lengths along the same tethered DNA molecule. We observed that FtsK assembled stepwise on DNA into a single hexamer, and began translocation rapidly (∼ 0.25 s). Without extruding DNA loops, single FtsK hexamers approached XerCD-dif and resided there for ∼ 0.5 s irrespective of whether XerCD-dif was synapsed or unsynapsed. FtsK then dissociated, rather than reversing. Infrequently, FtsK activated XerCD-dif recombination when it encountered a preformed synaptic complex, and dissociated before the completion of recombination, consistent with each FtsK-XerCD-dif encounter activating only one round of recombination.

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Citations

Jun 19, 2016·Proceedings of the National Academy of Sciences of the United States of America·Florian FournesPhilippe Rousseau
Jul 17, 2016·Proceedings of the National Academy of Sciences of the United States of America·Caroline Midonet, François-Xavier Barre
Aug 20, 2019·Proceedings of the National Academy of Sciences of the United States of America·Caroline MidonetFrançois-Xavier Barre
Apr 5, 2017·Frontiers in Microbiology·Fabio CastilloGeorge Szatmari
Jul 25, 2018·Proceedings of the National Academy of Sciences of the United States of America·Matthias SchickingerHendrik Dietz
Jun 14, 2019·Heredity·David Sherratt
Sep 3, 2019·Current Biology : CB·Philipp M WeberSilvia Bulgheresi

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