Cell cycle-dependent processing of DNA lesions controls localization of Rad9 to sites of genotoxic stress
Open accessAbstract
The Rad9/Rad1/Hus1 complex functions to facilitate the ATR-mediated phosphorylation of several substrates that control the checkpoint arrest induced by DNA damage. Here we show that in response to genotoxic stress induced by different types of damaging agents, Rad9 rapidly relocalized to sites of single stranded DNA, as visualized by discrete nuclear foci that co-localize with RPA. UV light-induced Rad9 foci also colocalized with TopBP1 and gamma-H2AX. Interestingly, Rad9 foci were predominately formed in G(1) and S phase after UV light, while treatment of cells with ionizing radiation (IR) resulted in accumulation of Rad9 into foci in S and G(2). Photobleaching experiments in living cells revealed that the Rad9 protein is highly mobile in undamaged cells. However, genotoxic stress induced the immobilization of a large proportion of the protein. The proportion of Rad9 immobilization was larger in S phase and the accumulation to sites of locally damaged areas induced by UV-laser irradiation was faster during DNA replication. Inactivation of nucleotide excision repair by knock down of XPA and XPC resulted in a decrease of G(1) phase cells that displayed Rad9 foci in response to UV light, whereas IR-induced Rad9 foci were not affected. In contrast, downregulation of CtIP, which promotes DSB resection, abrogated the IR-induced Rad9 foci. These findings show that due to processing of DNA lesions into a common intermediate, which occurs in a cell cycle-dependent manner, Rad9 is able to respond to different types of genotoxic stress.
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In human G1 cells, recruitment of the 9-1-1 complex to damaged DNA depends on XPA and XPC.
“In human G1 cells the recruitment of the 9-1-1 complex onto damaged DNA is dependent on XPA and XPC proteins [92]”
From Mind the gap: keeping UV lesions in check (2011) · cites this paper as [92]
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- Authors
- Warmerdam, D. O., Freire, R., Kanaar, R., & Smits, V. A. J.
- Affiliations
- Erasmus MC (NL)
- Hospital Universitario de Canarias (ES)
- Corresponding author
- Smits, Veronique A.J.
- Venue
- Cell Cycle
- Publisher
- Informa UK Limited
- Published
- 2009-06
- Volume
- 8
- Issue
- 11
- Pages
- 1765-1774
- ISSN
- 1538-4101 (ISSN-L)
- 1551-4005
- OpenAlex
- W2007795814
- Cited by
- 28
- References
- 0
- Citations by year
- 2026: 1 · 2019: 1 · 2017: 1 · 2016: 1 · 2014: 1 · 2013: 6 · 2012: 4
- FWCI
- 2.28
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Cell Cycle (publishedVersion)
- PubMed (publishedVersion)
- Pure Amsterdam UMC (publishedVersion, other-oa)
- EUR Research Repository (Erasmus University Rotterdam) (submittedVersion)
- Funders
- European Commission (512113, EMCR 2005-3412)
- Ministerio de Ciencia e Innovación (512113, EMCR 2005-3412)
- Instituto de Salud Carlos III
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Keywords
- Rad9, DNA damage response, ATR signaling, single-stranded DNA, RPA, TopBP1, γH2AX, UV-induced DNA damage, ionizing radiation, DNA replication, protein immobilization, nucleotide excision repair, XPA, XPC, CtIP, DNA double-strand break resection
- MeSH terms
- Carrier Proteins, Cell Cycle, DNA Damage, DNA-Binding Proteins, DNA, Single-Stranded, Endodeoxyribonucleases, HeLa Cells, Histones, Humans, Infrared Rays, Nuclear Proteins, Ultraviolet Rays, G1 Phase, S Phase, Protein Serine-Threonine Kinases, Cell Cycle Proteins, Genomic Instability, Cell Line, Tumor, Replication Protein A, Xeroderma Pigmentosum Group A Protein, Ataxia Telangiectasia Mutated Proteins
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