Two mechanisms coordinate replication termination by the Escherichia coli Tus–Ter complex

Handle URI:
http://hdl.handle.net/10754/579489
Title:
Two mechanisms coordinate replication termination by the Escherichia coli Tus–Ter complex
Authors:
Pandey, Manjula; Elshenawy, Mohamed ( 0000-0002-8599-8388 ) ; Jergic, Slobodan; Takahashi, Masateru; Dixon, Nicholas E.; Hamdan, Samir ( 0000-0001-5192-1852 ) ; Patel, Smita S.
Abstract:
The Escherichia coli replication terminator protein (Tus) binds to Ter sequences to block replication forks approaching from one direction. Here, we used single molecule and transient state kinetics to study responses of the heterologous phage T7 replisome to the Tus–Ter complex. The T7 replisome was arrested at the non-permissive end of Tus–Ter in a manner that is explained by a composite mousetrap and dynamic clamp model. An unpaired C(6) that forms a lock by binding into the cytosine binding pocket of Tus was most effective in arresting the replisome and mutation of C(6) removed the barrier. Isolated helicase was also blocked at the non-permissive end, but unexpectedly the isolated polymerase was not, unless C(6) was unpaired. Instead, the polymerase was blocked at the permissive end. This indicates that the Tus–Ter mechanism is sensitive to the translocation polarity of the DNA motor. The polymerase tracking along the template strand traps the C(6) to prevent lock formation; the helicase tracking along the other strand traps the complementary G(6) to aid lock formation. Our results are consistent with the model where strand separation by the helicase unpairs the GC(6) base pair and triggers lock formation immediately before the polymerase can sequester the C(6) base.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division
Citation:
Two mechanisms coordinate replication termination by the Escherichia coli Tus–Ter complex 2015, 43 (12):5924 Nucleic Acids Research
Publisher:
Oxford University Press (OUP)
Journal:
Nucleic Acids Research
Issue Date:
13-Jul-2015
DOI:
10.1093/nar/gkv527
Type:
Article
ISSN:
0305-1048; 1362-4962
Additional Links:
http://nar.oxfordjournals.org/lookup/doi/10.1093/nar/gkv527
Appears in Collections:
Articles; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorPandey, Manjulaen
dc.contributor.authorElshenawy, Mohameden
dc.contributor.authorJergic, Slobodanen
dc.contributor.authorTakahashi, Masateruen
dc.contributor.authorDixon, Nicholas E.en
dc.contributor.authorHamdan, Samiren
dc.contributor.authorPatel, Smita S.en
dc.date.accessioned2015-10-08T07:40:33Zen
dc.date.available2015-10-08T07:40:33Zen
dc.date.issued2015-07-13en
dc.identifier.citationTwo mechanisms coordinate replication termination by the Escherichia coli Tus–Ter complex 2015, 43 (12):5924 Nucleic Acids Researchen
dc.identifier.issn0305-1048en
dc.identifier.issn1362-4962en
dc.identifier.doi10.1093/nar/gkv527en
dc.identifier.urihttp://hdl.handle.net/10754/579489en
dc.description.abstractThe Escherichia coli replication terminator protein (Tus) binds to Ter sequences to block replication forks approaching from one direction. Here, we used single molecule and transient state kinetics to study responses of the heterologous phage T7 replisome to the Tus–Ter complex. The T7 replisome was arrested at the non-permissive end of Tus–Ter in a manner that is explained by a composite mousetrap and dynamic clamp model. An unpaired C(6) that forms a lock by binding into the cytosine binding pocket of Tus was most effective in arresting the replisome and mutation of C(6) removed the barrier. Isolated helicase was also blocked at the non-permissive end, but unexpectedly the isolated polymerase was not, unless C(6) was unpaired. Instead, the polymerase was blocked at the permissive end. This indicates that the Tus–Ter mechanism is sensitive to the translocation polarity of the DNA motor. The polymerase tracking along the template strand traps the C(6) to prevent lock formation; the helicase tracking along the other strand traps the complementary G(6) to aid lock formation. Our results are consistent with the model where strand separation by the helicase unpairs the GC(6) base pair and triggers lock formation immediately before the polymerase can sequester the C(6) base.en
dc.language.isoenen
dc.publisherOxford University Press (OUP)en
dc.relation.urlhttp://nar.oxfordjournals.org/lookup/doi/10.1093/nar/gkv527en
dc.rightsThis is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited.en
dc.titleTwo mechanisms coordinate replication termination by the Escherichia coli Tus–Ter complexen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.identifier.journalNucleic Acids Researchen
dc.eprint.versionPublisher's Version/PDFen
dc.contributor.institutionDepartment of Biochemistry and Molecular Biology, Rutgers, the State University of New Jersey, Robert Wood Johnson Medical School, Piscataway, NJ 08854, USAen
dc.contributor.institutionCentre for Medical and Molecular Bioscience, University of Wollongong, New South Wales 2522, Australiaen
dc.contributor.affiliationKing Abdullah University of Science and Technology (KAUST)en
kaust.authorHamdan, Samiren
kaust.authorElshenawy, Mohameden
kaust.authorTakahashi, Masateruen
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