The Rad4(TopBP1) ATR-activation Domain Functions in G1/S Phase in a Chromatin-dependent Manner

Overview

Journal PLoS Genet

Specialty Genetics

Date 2012 Jul 5

PMID 22761595

Citations 17

Authors

Su-Jiun Lin

Christopher P Wardlaw

Takashi Morishita

Izumi Miyabe

Charly Chahwan

Thomas Caspari

Ulrike Schmidt

Antony M Carr

Valerie Garcia

Affiliations

Soon will be listed here.

Abstract

DNA damage checkpoint activation can be subdivided in two steps: initial activation and signal amplification. The events distinguishing these two phases and their genetic determinants remain obscure. TopBP1, a mediator protein containing multiple BRCT domains, binds to and activates the ATR/ATRIP complex through its ATR-Activation Domain (AAD). We show that Schizosaccharomyces pombe Rad4(TopBP1) AAD-defective strains are DNA damage sensitive during G1/S-phase, but not during G2. Using lacO-LacI tethering, we developed a DNA damage-independent assay for checkpoint activation that is Rad4(TopBP1) AAD-dependent. In this assay, checkpoint activation requires histone H2A phosphorylation, the interaction between TopBP1 and the 9-1-1 complex, and is mediated by the phospho-binding activity of Crb2(53BP1). Consistent with a model where Rad4(TopBP1) AAD-dependent checkpoint activation is ssDNA/RPA-independent and functions to amplify otherwise weak checkpoint signals, we demonstrate that the Rad4(TopBP1) AAD is important for Chk1 phosphorylation when resection is limited in G2 by ablation of the resecting nuclease, Exo1. We also show that the Rad4(TopBP1) AAD acts additively with a Rad9 AAD in G1/S phase but not G2. We propose that AAD-dependent Rad3(ATR) checkpoint amplification is particularly important when DNA resection is limiting. In S. pombe, this manifests in G1/S phase and relies on protein-chromatin interactions.

Citing Articles

ATR-binding lncRNA ScaRNA2 promotes cancer resistance through facilitating efficient DNA end resection during homologous recombination repair.

Chen Y, Shen H, Liu T, Cao K, Wan Z, Du Z J Exp Clin Cancer Res. 2023; 42(1):256.

PMID: 37775817 PMC: 10542231. DOI: 10.1186/s13046-023-02829-4.

Phosphorylation-dependent assembly of DNA damage response systems and the central roles of TOPBP1.

Day M, Oliver A, Pearl L DNA Repair (Amst). 2021; 108:103232.

PMID: 34678589 PMC: 8651625. DOI: 10.1016/j.dnarep.2021.103232.

Activation of ATR-related protein kinase upon DNA damage recognition.

Ma M, Rodriguez A, Sugimoto K Curr Genet. 2019; 66(2):327-333.

PMID: 31624858 PMC: 7073305. DOI: 10.1007/s00294-019-01039-w.

BRCT domains of the DNA damage checkpoint proteins TOPBP1/Rad4 display distinct specificities for phosphopeptide ligands.

Day M, Rappas M, Ptasinska K, Boos D, Oliver A, Pearl L Elife. 2018; 7.

PMID: 30295604 PMC: 6175577. DOI: 10.7554/eLife.39979.

ATR-mediated proteome remodeling is a major determinant of homologous recombination capacity in cancer cells.

Kim D, Liu Y, Oberly S, Freire R, Smolka M Nucleic Acids Res. 2018; 46(16):8311-8325.

PMID: 30010936 PMC: 6144784. DOI: 10.1093/nar/gky625.

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