Chk1 loss creates replication barriers that compromise cell survival independently of excess origin firing.
Cell Line, Tumor
Cell Proliferation
Cell Survival
Checkpoint Kinase 1
/ genetics
DNA Damage
DNA Replication
DNA-Directed DNA Polymerase
/ metabolism
Gene Expression Regulation, Neoplastic
Gene Knockdown Techniques
/ methods
HCT116 Cells
HEK293 Cells
Humans
Neoplasms
/ genetics
Phosphorylation
Replication Origin
checkpoint kinase 1
origin firing
replication barrier
replicative helicase
translesion DNA polymerase eta
Journal
The EMBO journal
ISSN: 1460-2075
Titre abrégé: EMBO J
Pays: England
ID NLM: 8208664
Informations de publication
Date de publication:
15 08 2019
15 08 2019
Historique:
received:
03
12
2018
revised:
11
06
2019
accepted:
19
06
2019
pubmed:
12
7
2019
medline:
24
12
2019
entrez:
12
7
2019
Statut:
ppublish
Résumé
The effectiveness of checkpoint kinase 1 (Chk1) inhibitors at killing cancer cells is considered to be fully dependent on their effect on DNA replication initiation. Chk1 inhibition boosts origin firing, presumably limiting the availability of nucleotides and in turn provoking the slowdown and subsequent collapse of forks, thus decreasing cell viability. Here we show that slow fork progression in Chk1-inhibited cells is not an indirect effect of excess new origin firing. Instead, fork slowdown results from the accumulation of replication barriers, whose bypass is impeded by CDK-dependent phosphorylation of the specialized DNA polymerase eta (Polη). Also in contrast to the linear model, the accumulation of DNA damage in Chk1-deficient cells depends on origin density but is largely independent of fork speed. Notwithstanding this, origin dysregulation contributes only mildly to the poor proliferation rates of Chk1-depleted cells. Moreover, elimination of replication barriers by downregulation of helicase components, but not their bypass by Polη, improves cell survival. Our results thus shed light on the molecular basis of the sensitivity of tumors to Chk1 inhibition.
Identifiants
pubmed: 31294866
doi: 10.15252/embj.2018101284
pmc: PMC6694221
doi:
Substances chimiques
CHEK1 protein, human
EC 2.7.11.1
Checkpoint Kinase 1
EC 2.7.11.1
DNA-Directed DNA Polymerase
EC 2.7.7.7
Rad30 protein
EC 2.7.7.7
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e101284Subventions
Organisme : MINCyT|Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT)
ID : PICT2015-1217
Pays : International
Organisme : MINCyT|Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT)
ID : PICT2014-0522
Pays : International
Organisme : MINCyT|Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT)
ID : PICT2016-1239
Pays : International
Organisme : Institut National Du Cancer (INCa)
Pays : International
Organisme : Agence Nationale de la Recherche (ANR)
Pays : International
Organisme : Associazione Italiana per la Ricerca sul Cancro (AIRC)
ID : 12710
Pays : International
Organisme : European Commission (EC)
ID : PCIG10-GA-2011-303806
Pays : International
Informations de copyright
© 2019 The Authors.
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