Pif1-family helicases support fork convergence during DNA replication termination in eukaryotes

Thomas Deegan (Lead / Corresponding author), Jonathan Baxter, Maria Ortiz Bazan, Joseph T. P. Yeeles, Karim Labib (Lead / Corresponding author)

Research output: Contribution to journalArticle

7 Citations (Scopus)
79 Downloads (Pure)

Abstract

The convergence of two DNA replication forks creates unique problems during DNA replication termination. In E. coli and SV40, the release of torsional strain by type II topoisomerases is critical for converging replisomes to complete DNA synthesis, but the pathways that mediate fork convergence in eukaryotes are unknown. We studied the convergence of reconstituted yeast replication forks that include all core replisome components and both type I and type II topoisomerases. We found that most converging forks stall at a very late stage, indicating a role for additional factors. We showed that the Pif1 and Rrm3 DNA helicases promote efficient fork convergence and completion of DNA synthesis, even in the absence of type II topoisomerase. Furthermore, Rrm3 and Pif1 are also important for termination of plasmid DNA replication in vivo. These findings identify a eukaryotic pathway for DNA replication termination that is distinct from previously characterized prokaryotic mechanisms.

Original languageEnglish
Pages (from-to)231-244.e9
Number of pages24
JournalMolecular Cell
Volume74
Issue number2
Early online date5 Mar 2019
DOIs
Publication statusPublished - 18 Apr 2019

Fingerprint

Eukaryota
DNA Replication
Type II DNA Topoisomerase
DNA Helicases
Type I DNA Topoisomerase
DNA
Plasmids
Yeasts
Escherichia coli

Keywords

  • CMG helicase
  • DNA replication termination
  • Pif1
  • Rrm3
  • Top2
  • chromosome replication
  • fork convergence
  • replisome
  • topoisomerase

Cite this

Deegan, Thomas ; Baxter, Jonathan ; Ortiz Bazan, Maria ; Yeeles, Joseph T. P. ; Labib, Karim. / Pif1-family helicases support fork convergence during DNA replication termination in eukaryotes. In: Molecular Cell. 2019 ; Vol. 74, No. 2. pp. 231-244.e9.
@article{12002532f1cc475587c94f76b4a1aa91,
title = "Pif1-family helicases support fork convergence during DNA replication termination in eukaryotes",
abstract = "The convergence of two DNA replication forks creates unique problems during DNA replication termination. In E. coli and SV40, the release of torsional strain by type II topoisomerases is critical for converging replisomes to complete DNA synthesis, but the pathways that mediate fork convergence in eukaryotes are unknown. We studied the convergence of reconstituted yeast replication forks that include all core replisome components and both type I and type II topoisomerases. We found that most converging forks stall at a very late stage, indicating a role for additional factors. We showed that the Pif1 and Rrm3 DNA helicases promote efficient fork convergence and completion of DNA synthesis, even in the absence of type II topoisomerase. Furthermore, Rrm3 and Pif1 are also important for termination of plasmid DNA replication in vivo. These findings identify a eukaryotic pathway for DNA replication termination that is distinct from previously characterized prokaryotic mechanisms.",
keywords = "CMG helicase, DNA replication termination, Pif1, Rrm3, Top2, chromosome replication, fork convergence, replisome, topoisomerase",
author = "Thomas Deegan and Jonathan Baxter and {Ortiz Bazan}, Maria and Yeeles, {Joseph T. P.} and Karim Labib",
note = "We gratefully acknowledge the support of the Wellcome Trust (reference 204678/Z/16/Z for a Sir Henry Wellcome Postdoctoral Fellowship to TD and 102943/Z/13/Z for an Investigator award to KL), the Medical Research Council (core grants MC_UU_12016/13 to KL and MC_UP_1201/12 to JY), BBSRC (project grant BB/N007344/1 to JB) and the Royal Society (Research Fellowship to JB).",
year = "2019",
month = "4",
day = "18",
doi = "10.1016/j.molcel.2019.01.040",
language = "English",
volume = "74",
pages = "231--244.e9",
journal = "Molecular Cell",
issn = "1097-2765",
publisher = "Elsevier",
number = "2",

}

Pif1-family helicases support fork convergence during DNA replication termination in eukaryotes. / Deegan, Thomas (Lead / Corresponding author); Baxter, Jonathan; Ortiz Bazan, Maria; Yeeles, Joseph T. P.; Labib, Karim (Lead / Corresponding author).

In: Molecular Cell, Vol. 74, No. 2, 18.04.2019, p. 231-244.e9.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Pif1-family helicases support fork convergence during DNA replication termination in eukaryotes

AU - Deegan, Thomas

AU - Baxter, Jonathan

AU - Ortiz Bazan, Maria

AU - Yeeles, Joseph T. P.

AU - Labib, Karim

N1 - We gratefully acknowledge the support of the Wellcome Trust (reference 204678/Z/16/Z for a Sir Henry Wellcome Postdoctoral Fellowship to TD and 102943/Z/13/Z for an Investigator award to KL), the Medical Research Council (core grants MC_UU_12016/13 to KL and MC_UP_1201/12 to JY), BBSRC (project grant BB/N007344/1 to JB) and the Royal Society (Research Fellowship to JB).

PY - 2019/4/18

Y1 - 2019/4/18

N2 - The convergence of two DNA replication forks creates unique problems during DNA replication termination. In E. coli and SV40, the release of torsional strain by type II topoisomerases is critical for converging replisomes to complete DNA synthesis, but the pathways that mediate fork convergence in eukaryotes are unknown. We studied the convergence of reconstituted yeast replication forks that include all core replisome components and both type I and type II topoisomerases. We found that most converging forks stall at a very late stage, indicating a role for additional factors. We showed that the Pif1 and Rrm3 DNA helicases promote efficient fork convergence and completion of DNA synthesis, even in the absence of type II topoisomerase. Furthermore, Rrm3 and Pif1 are also important for termination of plasmid DNA replication in vivo. These findings identify a eukaryotic pathway for DNA replication termination that is distinct from previously characterized prokaryotic mechanisms.

AB - The convergence of two DNA replication forks creates unique problems during DNA replication termination. In E. coli and SV40, the release of torsional strain by type II topoisomerases is critical for converging replisomes to complete DNA synthesis, but the pathways that mediate fork convergence in eukaryotes are unknown. We studied the convergence of reconstituted yeast replication forks that include all core replisome components and both type I and type II topoisomerases. We found that most converging forks stall at a very late stage, indicating a role for additional factors. We showed that the Pif1 and Rrm3 DNA helicases promote efficient fork convergence and completion of DNA synthesis, even in the absence of type II topoisomerase. Furthermore, Rrm3 and Pif1 are also important for termination of plasmid DNA replication in vivo. These findings identify a eukaryotic pathway for DNA replication termination that is distinct from previously characterized prokaryotic mechanisms.

KW - CMG helicase

KW - DNA replication termination

KW - Pif1

KW - Rrm3

KW - Top2

KW - chromosome replication

KW - fork convergence

KW - replisome

KW - topoisomerase

UR - http://www.scopus.com/inward/record.url?scp=85064161387&partnerID=8YFLogxK

U2 - 10.1016/j.molcel.2019.01.040

DO - 10.1016/j.molcel.2019.01.040

M3 - Article

C2 - 30850330

VL - 74

SP - 231-244.e9

JO - Molecular Cell

JF - Molecular Cell

SN - 1097-2765

IS - 2

ER -