Delineation of the Ancestral Tus-Dependent Replication Fork Trap

Casey Toft; Morgane Moreau; Jiri Perutka; Savitri Mandapati; Peter Enyeart; Alanna Sorenson; Andrew D. Ellington; Patrick Schaeffer

doi:10.20944/preprints202111.0539.v1

Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Version 1 : Received: 24 November 2021 / Approved: 29 November 2021 / Online: 29 November 2021 (12:52:31 CET)

In Escherichia coli, DNA replication termination is orchestrated by two clusters of Ter sites forming a DNA replication fork trap when bound by Tus proteins. The formation of a ‘locked’ Tus-Ter complex is essential for halting incoming DNA replication forks. However, the absence of replication fork arrest at some Ter sites raised questions about their significance. In this study, we examined the genome-wide distribution of Tus and found that only the six innermost Ter sites (TerA-E and G) were significantly bound by Tus. We also found that a single ectopic insertion of TerB in its non-permissive orientation could not be achieved, advocating against a need for ‘back-up’ Ter sites. Finally, examination of the genomes of a variety of Enterobacterales revealed a new replication fork trap architecture mostly found outside the Enterobacteriaceae family. Taken together, our data enabled the delineation of a narrow ancestral Tus-dependent DNA replication fork trap consisting of only two Ter sites.

Replication fork trap; Tus-Ter; dif; ChIP-Seq; GC-skew; Enterobacterales

Biology and Life Sciences, Biochemistry and Molecular Biology

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Delineation of the Ancestral Tus-Dependent Replication Fork Trap

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