CRL2(Lrr1) promotes unloading of the vertebrate replisome from chromatin during replication termination

TY - JOUR

T1 - CRL2(Lrr1) promotes unloading of the vertebrate replisome from chromatin during replication termination

AU - Dewar, James M

AU - Low, Emily

AU - Mann, Matthias

AU - Räschle, Markus

AU - Walter, Johannes C

N1 - © 2017 Dewar et al.; Published by Cold Spring Harbor Laboratory Press.

PY - 2017/2/1

Y1 - 2017/2/1

N2 - A key event during eukaryotic replication termination is the removal of the CMG helicase from chromatin. CMG unloading involves ubiquitylation of its Mcm7 subunit and the action of the p97 ATPase. Using a proteomic screen in Xenopus egg extracts, we identified factors that are enriched on chromatin when CMG unloading is blocked. This approach identified the E3 ubiquitin ligase CRL2(Lrr1), a specific p97 complex, other potential regulators of termination, and many replisome components. We show that Mcm7 ubiquitylation and CRL2(Lrr1) binding to chromatin are temporally linked and occur only during replication termination. In the absence of CRL2(Lrr1), Mcm7 is not ubiquitylated, CMG unloading is inhibited, and a large subcomplex of the vertebrate replisome that includes DNA Pol ε is retained on DNA. Our data identify CRL2(Lrr1) as a master regulator of replisome disassembly during vertebrate DNA replication termination.

AB - A key event during eukaryotic replication termination is the removal of the CMG helicase from chromatin. CMG unloading involves ubiquitylation of its Mcm7 subunit and the action of the p97 ATPase. Using a proteomic screen in Xenopus egg extracts, we identified factors that are enriched on chromatin when CMG unloading is blocked. This approach identified the E3 ubiquitin ligase CRL2(Lrr1), a specific p97 complex, other potential regulators of termination, and many replisome components. We show that Mcm7 ubiquitylation and CRL2(Lrr1) binding to chromatin are temporally linked and occur only during replication termination. In the absence of CRL2(Lrr1), Mcm7 is not ubiquitylated, CMG unloading is inhibited, and a large subcomplex of the vertebrate replisome that includes DNA Pol ε is retained on DNA. Our data identify CRL2(Lrr1) as a master regulator of replisome disassembly during vertebrate DNA replication termination.

KW - Adenosine Triphosphatases

KW - Animals

KW - Chromatin

KW - DNA

KW - DNA Helicases

KW - DNA Polymerase II

KW - DNA Replication

KW - Minichromosome Maintenance Complex Component 7

KW - Nuclear Proteins

KW - Ubiquitin-Protein Ligases

KW - Ubiquitination

KW - Xenopus Proteins

KW - Xenopus laevis

KW - Journal Article

U2 - 10.1101/gad.291799.116

DO - 10.1101/gad.291799.116

M3 - Journal article

C2 - 28235849

SN - 0890-9369

VL - 31

SP - 275

EP - 290

JO - Genes & Development

JF - Genes & Development

IS - 3

ER -