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

Stefano Di Marco; Zdenka Hasanova; Radhakrishnan Kanagaraj; Nagaraja Chappidi; Veronika Altmannova; Shruti Menon; Hana Sedlackova; Jana Langhoff; Kalpana Surendranath; Daniela Hühn; Rahul Bhowmick; Victoria Marini; Stefano Ferrari; Ian D Hickson; Lumir Krejci; Pavel Janscak

doi:10.1016/j.molcel.2017.05.006

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

Stefano Di Marco, Zdenka Hasanova, Radhakrishnan Kanagaraj, Nagaraja Chappidi, Veronika Altmannova, Shruti Menon, Hana Sedlackova, Jana Langhoff, Kalpana Surendranath, Daniela Hühn, Rahul Bhowmick, Victoria Marini, Stefano Ferrari, Ian D Hickson, Lumir Krejci, Pavel Janscak

40 Citations (Scopus)

Abstract

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.

Original language	English
Journal	Molecular Cell
Volume	66
Issue number	5
Pages (from-to)	658-671.e8
ISSN	1097-2765
DOIs	https://doi.org/10.1016/j.molcel.2017.05.006
Publication status	Published - 1 Jun 2017

Keywords

Binding Sites
Chromosomal Instability
Chromosome Fragile Sites
Chromosome Segregation
Cyclin-Dependent Kinases
DNA
DNA Damage
DNA Repair
DNA-Binding Proteins
Endodeoxyribonucleases
Endonucleases
HEK293 Cells
HeLa Cells
Humans
Mitosis
Phosphorylation
Protein Binding
RNA Interference
Rad51 Recombinase
RecQ Helicases
Replication Origin
Time Factors
Transfection
Journal Article

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10.1016/j.molcel.2017.05.006

http://www.cell.com/article/S1097276517303192/pdf

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Di Marco, S., Hasanova, Z., Kanagaraj, R., Chappidi, N., Altmannova, V., Menon, S., Sedlackova, H., Langhoff, J., Surendranath, K., Hühn, D., Bhowmick, R., Marini, V., Ferrari, S., Hickson, I. D., Krejci, L., & Janscak, P. (2017). RECQ5 Helicase Cooperates with MUS81 Endonuclease in Processing Stalled Replication Forks at Common Fragile Sites during Mitosis. Molecular Cell, 66(5), 658-671.e8. https://doi.org/10.1016/j.molcel.2017.05.006

Di Marco, S, Hasanova, Z, Kanagaraj, R, Chappidi, N, Altmannova, V, Menon, S, Sedlackova, H, Langhoff, J, Surendranath, K, Hühn, D, Bhowmick, R, Marini, V, Ferrari, S, Hickson, ID, Krejci, L & Janscak, P 2017, 'RECQ5 Helicase Cooperates with MUS81 Endonuclease in Processing Stalled Replication Forks at Common Fragile Sites during Mitosis', Molecular Cell, vol. 66, no. 5, pp. 658-671.e8. https://doi.org/10.1016/j.molcel.2017.05.006

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

abstract = "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.",

keywords = "Binding Sites, Chromosomal Instability, Chromosome Fragile Sites, Chromosome Segregation, Cyclin-Dependent Kinases, DNA, DNA Damage, DNA Repair, DNA-Binding Proteins, Endodeoxyribonucleases, Endonucleases, HEK293 Cells, HeLa Cells, Humans, Mitosis, Phosphorylation, Protein Binding, RNA Interference, Rad51 Recombinase, RecQ Helicases, Replication Origin, Time Factors, Transfection, Journal Article",

author = "{Di Marco}, Stefano and Zdenka Hasanova and Radhakrishnan Kanagaraj and Nagaraja Chappidi and Veronika Altmannova and Shruti Menon and Hana Sedlackova and Jana Langhoff and Kalpana Surendranath and Daniela H{\"u}hn and Rahul Bhowmick and Victoria Marini and Stefano Ferrari and Hickson, {Ian D} and Lumir Krejci and Pavel Janscak",

year = "2017",

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doi = "10.1016/j.molcel.2017.05.006",

language = "English",

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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

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 -

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

Abstract

Keywords

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