RECQ5 Helicase Cooperates with MUS81 Endonuclease in Processing Stalled Replication Forks at Common Fragile Sites during Mitosis

TY - JOUR

T1 - RECQ5 Helicase Cooperates with MUS81 Endonuclease in Processing Stalled Replication Forks at Common Fragile Sites during Mitosis

AU - Di Marco, Stefano

AU - Hasanova, Zdenka

AU - Kanagaraj, Radhakrishnan

AU - Chappidi, Nagaraja

AU - Altmannova, Veronika

AU - Menon, Shruti

AU - Sedlackova, Hana

AU - Langhoff, Jana

AU - Surendranath, Kalpana

AU - Hühn, Daniela

AU - Bhowmick, Rahul

AU - Marini, Victoria

AU - Ferrari, Stefano

AU - Hickson, Ian D

AU - Krejci, Lumir

AU - Janscak, Pavel

N1 - Copyright © 2017 Elsevier Inc. All rights reserved.

PY - 2017/6/1

Y1 - 2017/6/1

N2 - The MUS81-EME1 endonuclease cleaves late replication intermediates at common fragile sites (CFSs) during early mitosis to trigger DNA-repair synthesis that ensures faithful chromosome segregation. Here, we show that these DNA transactions are promoted by RECQ5 DNA helicase in a manner dependent on its Ser727 phosphorylation by CDK1. Upon replication stress, RECQ5 associates with CFSs in early mitosis through its physical interaction with MUS81 and promotes MUS81-dependent mitotic DNA synthesis. RECQ5 depletion or mutational inactivation of its ATP-binding site, RAD51-interacting domain, or phosphorylation site causes excessive binding of RAD51 to CFS loci and impairs CFS expression. This leads to defective chromosome segregation and accumulation of CFS-associated DNA damage in G1 cells. Biochemically, RECQ5 alleviates the inhibitory effect of RAD51 on 3'-flap DNA cleavage by MUS81-EME1 through its RAD51 filament disruption activity. These data suggest that RECQ5 removes RAD51 filaments stabilizing stalled replication forks at CFSs and hence facilitates CFS cleavage by MUS81-EME1.

AB - The MUS81-EME1 endonuclease cleaves late replication intermediates at common fragile sites (CFSs) during early mitosis to trigger DNA-repair synthesis that ensures faithful chromosome segregation. Here, we show that these DNA transactions are promoted by RECQ5 DNA helicase in a manner dependent on its Ser727 phosphorylation by CDK1. Upon replication stress, RECQ5 associates with CFSs in early mitosis through its physical interaction with MUS81 and promotes MUS81-dependent mitotic DNA synthesis. RECQ5 depletion or mutational inactivation of its ATP-binding site, RAD51-interacting domain, or phosphorylation site causes excessive binding of RAD51 to CFS loci and impairs CFS expression. This leads to defective chromosome segregation and accumulation of CFS-associated DNA damage in G1 cells. Biochemically, RECQ5 alleviates the inhibitory effect of RAD51 on 3'-flap DNA cleavage by MUS81-EME1 through its RAD51 filament disruption activity. These data suggest that RECQ5 removes RAD51 filaments stabilizing stalled replication forks at CFSs and hence facilitates CFS cleavage by MUS81-EME1.

KW - Binding Sites

KW - Chromosomal Instability

KW - Chromosome Fragile Sites

KW - Chromosome Segregation

KW - Cyclin-Dependent Kinases

KW - DNA

KW - DNA Damage

KW - DNA Repair

KW - DNA-Binding Proteins

KW - Endodeoxyribonucleases

KW - Endonucleases

KW - HEK293 Cells

KW - HeLa Cells

KW - Humans

KW - Mitosis

KW - Phosphorylation

KW - Protein Binding

KW - RNA Interference

KW - Rad51 Recombinase

KW - RecQ Helicases

KW - Replication Origin

KW - Time Factors

KW - Transfection

KW - Journal Article

U2 - 10.1016/j.molcel.2017.05.006

DO - 10.1016/j.molcel.2017.05.006

M3 - Journal article

C2 - 28575661

SN - 1097-2765

VL - 66

SP - 658-671.e8

JO - Molecular Cell

JF - Molecular Cell

IS - 5

ER -