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Exchange between Escherichia coli polymerases II and III on a processivity clamp

Escherichia coli has three DNA polymerases implicated in the bypass of DNA damage, a process called translesion synthesis (TLS) that alleviates replication stalling. Although these polymerases are specialized for different DNA lesions, it is unclear if they interact differently with the replication...

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Autores principales: Kath, James E., Chang, Seungwoo, Scotland, Michelle K., Wilbertz, Johannes H., Jergic, Slobodan, Dixon, Nicholas E., Sutton, Mark D., Loparo, Joseph J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4770218/
https://www.ncbi.nlm.nih.gov/pubmed/26657641
http://dx.doi.org/10.1093/nar/gkv1375
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author Kath, James E.
Chang, Seungwoo
Scotland, Michelle K.
Wilbertz, Johannes H.
Jergic, Slobodan
Dixon, Nicholas E.
Sutton, Mark D.
Loparo, Joseph J.
author_facet Kath, James E.
Chang, Seungwoo
Scotland, Michelle K.
Wilbertz, Johannes H.
Jergic, Slobodan
Dixon, Nicholas E.
Sutton, Mark D.
Loparo, Joseph J.
author_sort Kath, James E.
collection PubMed
description Escherichia coli has three DNA polymerases implicated in the bypass of DNA damage, a process called translesion synthesis (TLS) that alleviates replication stalling. Although these polymerases are specialized for different DNA lesions, it is unclear if they interact differently with the replication machinery. Of the three, DNA polymerase (Pol) II remains the most enigmatic. Here we report a stable ternary complex of Pol II, the replicative polymerase Pol III core complex and the dimeric processivity clamp, β. Single-molecule experiments reveal that the interactions of Pol II and Pol III with β allow for rapid exchange during DNA synthesis. As with another TLS polymerase, Pol IV, increasing concentrations of Pol II displace the Pol III core during DNA synthesis in a minimal reconstitution of primer extension. However, in contrast to Pol IV, Pol II is inefficient at disrupting rolling-circle synthesis by the fully reconstituted Pol III replisome. Together, these data suggest a β-mediated mechanism of exchange between Pol II and Pol III that occurs outside the replication fork.
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spelling pubmed-47702182016-02-29 Exchange between Escherichia coli polymerases II and III on a processivity clamp Kath, James E. Chang, Seungwoo Scotland, Michelle K. Wilbertz, Johannes H. Jergic, Slobodan Dixon, Nicholas E. Sutton, Mark D. Loparo, Joseph J. Nucleic Acids Res Genome Integrity, Repair and Replication Escherichia coli has three DNA polymerases implicated in the bypass of DNA damage, a process called translesion synthesis (TLS) that alleviates replication stalling. Although these polymerases are specialized for different DNA lesions, it is unclear if they interact differently with the replication machinery. Of the three, DNA polymerase (Pol) II remains the most enigmatic. Here we report a stable ternary complex of Pol II, the replicative polymerase Pol III core complex and the dimeric processivity clamp, β. Single-molecule experiments reveal that the interactions of Pol II and Pol III with β allow for rapid exchange during DNA synthesis. As with another TLS polymerase, Pol IV, increasing concentrations of Pol II displace the Pol III core during DNA synthesis in a minimal reconstitution of primer extension. However, in contrast to Pol IV, Pol II is inefficient at disrupting rolling-circle synthesis by the fully reconstituted Pol III replisome. Together, these data suggest a β-mediated mechanism of exchange between Pol II and Pol III that occurs outside the replication fork. Oxford University Press 2016-02-29 2015-12-10 /pmc/articles/PMC4770218/ /pubmed/26657641 http://dx.doi.org/10.1093/nar/gkv1375 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This 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. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Genome Integrity, Repair and Replication
Kath, James E.
Chang, Seungwoo
Scotland, Michelle K.
Wilbertz, Johannes H.
Jergic, Slobodan
Dixon, Nicholas E.
Sutton, Mark D.
Loparo, Joseph J.
Exchange between Escherichia coli polymerases II and III on a processivity clamp
title Exchange between Escherichia coli polymerases II and III on a processivity clamp
title_full Exchange between Escherichia coli polymerases II and III on a processivity clamp
title_fullStr Exchange between Escherichia coli polymerases II and III on a processivity clamp
title_full_unstemmed Exchange between Escherichia coli polymerases II and III on a processivity clamp
title_short Exchange between Escherichia coli polymerases II and III on a processivity clamp
title_sort exchange between escherichia coli polymerases ii and iii on a processivity clamp
topic Genome Integrity, Repair and Replication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4770218/
https://www.ncbi.nlm.nih.gov/pubmed/26657641
http://dx.doi.org/10.1093/nar/gkv1375
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