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A Molecular Mousetrap Determines Polarity of Termination of DNA Replication in E. coli

A Molecular Mousetrap Determines Polarity of Termination of DNA Replication in E. coli
A Molecular Mousetrap Determines Polarity of Termination of DNA Replication in E. coli
During chromosome synthesis in Escherichia coli, replication forks are blocked by Tus bound Ter sites on approach from one direction but not the other. To study the basis of this polarity, we measured the rates of dissociation of Tus from forked TerB oligonucleotides, such as would be produced by the replicative DnaB helicase at both the fork-blocking (nonpermissive) and permissive ends of the Ter site. Strand separation of a few nucleotides at the permissive end was sufficient to force rapid dissociation of Tus to allow fork progression. In contrast, strand separation extending to and including the strictly conserved G-C(6) base pair at the nonpermissive end led to formation of a stable locked complex. Lock formation specifically requires the cytosine residue, C(6). The crystal structure of the locked complex showed that C(6) moves 14 Å from its normal position to bind in a cytosine-specific pocket on the surface of Tus.
0092-8674
1309-1319
Mulcair, M.D.
115c9b8d-bcc3-4fce-8454-097c048e97c9
Schaeffer, P.M.
e2e01729-220a-4663-b2d4-165eb360df37
Oakley, A.J.
19a6d409-fa7b-4533-838b-bee9cad54abe
Cross, H.F.
2ff9d443-6448-4b19-8679-2a1d9573a5b7
Neylon, D.C.
ab4fb130-bf8c-40d3-8408-acea4f372b8b
Hill, T.M.
63701932-31d5-4154-907c-864b150fdeb8
Dixon, N.E.
510ea408-c47a-4f71-ae4c-e31b2cc9a004
Mulcair, M.D.
115c9b8d-bcc3-4fce-8454-097c048e97c9
Schaeffer, P.M.
e2e01729-220a-4663-b2d4-165eb360df37
Oakley, A.J.
19a6d409-fa7b-4533-838b-bee9cad54abe
Cross, H.F.
2ff9d443-6448-4b19-8679-2a1d9573a5b7
Neylon, D.C.
ab4fb130-bf8c-40d3-8408-acea4f372b8b
Hill, T.M.
63701932-31d5-4154-907c-864b150fdeb8
Dixon, N.E.
510ea408-c47a-4f71-ae4c-e31b2cc9a004

Mulcair, M.D., Schaeffer, P.M., Oakley, A.J., Cross, H.F., Neylon, D.C., Hill, T.M. and Dixon, N.E. (2006) A Molecular Mousetrap Determines Polarity of Termination of DNA Replication in E. coli. Cell, 125 (7), 1309-1319. (doi:10.1016/j.cell.2006.04.040).

Record type: Article

Abstract

During chromosome synthesis in Escherichia coli, replication forks are blocked by Tus bound Ter sites on approach from one direction but not the other. To study the basis of this polarity, we measured the rates of dissociation of Tus from forked TerB oligonucleotides, such as would be produced by the replicative DnaB helicase at both the fork-blocking (nonpermissive) and permissive ends of the Ter site. Strand separation of a few nucleotides at the permissive end was sufficient to force rapid dissociation of Tus to allow fork progression. In contrast, strand separation extending to and including the strictly conserved G-C(6) base pair at the nonpermissive end led to formation of a stable locked complex. Lock formation specifically requires the cytosine residue, C(6). The crystal structure of the locked complex showed that C(6) moves 14 Å from its normal position to bind in a cytosine-specific pocket on the surface of Tus.

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Submitted date: 19 December 2005
Published date: 30 June 2006

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Local EPrints ID: 40707
URI: http://eprints.soton.ac.uk/id/eprint/40707
ISSN: 0092-8674
PURE UUID: f2db24a7-efd3-4cff-b3c1-f55b83900a57

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Date deposited: 15 Feb 2007
Last modified: 15 Mar 2024 08:21

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Contributors

Author: M.D. Mulcair
Author: P.M. Schaeffer
Author: A.J. Oakley
Author: H.F. Cross
Author: D.C. Neylon
Author: T.M. Hill
Author: N.E. Dixon

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