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

Journal Article


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Abstract


  • 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.

Authors


  •   Kath, James E. (external author)
  •   Chang, Seungwoo (external author)
  •   Scotland, Michelle K. (external author)
  •   Wilbertz, Johannes H. (external author)
  •   Jergic, Slobodan
  •   Dixon, Nicholas E.
  •   Sutton, Mark D. (external author)
  •   Loparo, Joseph J. (external author)

Publication Date


  • 2016

Citation


  • Kath, J. E., Chang, S., Scotland, M. K., Wilbertz, J. H., Jergic, S., Dixon, N. E., Sutton, M. D. & Loparo, J. J. (2016). Exchange between Escherichia coli polymerases II and III on a processivity clamp. Nucleic Acids Research, 44 (4), 1681-1690.

Scopus Eid


  • 2-s2.0-84960483772

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=1669&context=ihmri

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/647

Has Global Citation Frequency


Number Of Pages


  • 9

Start Page


  • 1681

End Page


  • 1690

Volume


  • 44

Issue


  • 4

Place Of Publication


  • United Kingdom

Abstract


  • 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.

Authors


  •   Kath, James E. (external author)
  •   Chang, Seungwoo (external author)
  •   Scotland, Michelle K. (external author)
  •   Wilbertz, Johannes H. (external author)
  •   Jergic, Slobodan
  •   Dixon, Nicholas E.
  •   Sutton, Mark D. (external author)
  •   Loparo, Joseph J. (external author)

Publication Date


  • 2016

Citation


  • Kath, J. E., Chang, S., Scotland, M. K., Wilbertz, J. H., Jergic, S., Dixon, N. E., Sutton, M. D. & Loparo, J. J. (2016). Exchange between Escherichia coli polymerases II and III on a processivity clamp. Nucleic Acids Research, 44 (4), 1681-1690.

Scopus Eid


  • 2-s2.0-84960483772

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=1669&context=ihmri

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/647

Has Global Citation Frequency


Number Of Pages


  • 9

Start Page


  • 1681

End Page


  • 1690

Volume


  • 44

Issue


  • 4

Place Of Publication


  • United Kingdom