ATR activation is regulated by dimerization of ATR activating proteins.
Adaptor Proteins, Signal Transducing
/ metabolism
Antigens, Surface
/ metabolism
Ataxia Telangiectasia Mutated Proteins
/ genetics
Carrier Proteins
/ metabolism
Cell Cycle Proteins
/ metabolism
Cell Line, Tumor
DNA Damage
/ genetics
DNA Repair
/ genetics
DNA Replication
/ genetics
DNA-Binding Proteins
/ metabolism
Dimerization
Humans
Nuclear Proteins
/ metabolism
Phosphorylation
Protein Binding
Protein Domains
/ genetics
Signal Transduction
/ physiology
ATR
ATR activation domain
DNA damage response
ETAA1
RPA
TOPBP1
cell cycle checkpoint
phosphatidylinositol 3-kinase related protein kinase
replication stress response
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
Historique:
received:
07
12
2020
revised:
17
02
2021
accepted:
22
02
2021
pubmed:
27
2
2021
medline:
31
8
2021
entrez:
26
2
2021
Statut:
ppublish
Résumé
The checkpoint kinase ATR regulates DNA repair, cell cycle progression, and other DNA damage and replication stress responses. ATR signaling is stimulated by an ATR activating protein, and in metazoan cells, there are at least two ATR activators: TOPBP1 and ETAA1. Current evidence indicates TOPBP1 and ETAA1 activate ATR via the same biochemical mechanism, but several aspects of this mechanism remain undefined. For example, ATR and its obligate binding partner ATR interacting protein (ATRIP) form a tetrameric complex consisting of two ATR and two ATRIP molecules, but whether TOPBP1 or ETAA1 dimerization is similarly required for ATR function is unclear. Here, we show that fusion of the TOPBP1 and ETAA1 ATR activation domains (AADs) to dimeric tags makes them more potent activators of ATR in vitro. Furthermore, induced dimerization of both AADs using chemical dimerization of a modified FKBP tag enhances ATR kinase activation and signaling in cells. ETAA1 forms oligomeric complexes mediated by regions of the protein that are predicted to be intrinsically disordered. Induced dimerization of a "mini-ETAA1" protein that contains the AAD and Replication Protein A (RPA) interaction motifs enhances ATR signaling, rescues cellular hypersensitivity to DNA damaging agents, and suppresses micronuclei formation in ETAA1-deficient cells. Together, our results indicate that TOPBP1 and ETAA1 dimerization is important for optimal ATR signaling and genome stability.
Identifiants
pubmed: 33636182
pii: S0021-9258(21)00228-3
doi: 10.1016/j.jbc.2021.100455
pmc: PMC7994790
pii:
doi:
Substances chimiques
ATRIP protein, human
0
Adaptor Proteins, Signal Transducing
0
Antigens, Surface
0
Carrier Proteins
0
Cell Cycle Proteins
0
DNA-Binding Proteins
0
ETAA1 protein, human
0
Nuclear Proteins
0
TOPBP1 protein, human
0
ATR protein, human
EC 2.7.11.1
Ataxia Telangiectasia Mutated Proteins
EC 2.7.11.1
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
100455Subventions
Organisme : NCI NIH HHS
ID : P50 CA098131
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA239161
Pays : United States
Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.
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