UV-induced photolesions elicit ATR-kinase-dependent signaling in non-cycling cells through nucleotide excision repair-dependent and -independent pathways

Vrouwe, Mischa G.; Pines, Alex; Overmeer, René M.; Hanada, Katsuhiro; Mullenders, Leon H.F.

doi:10.1242/jcs.075325

Cited by 95 publications

(93 citation statements)

References 64 publications

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“…We used GGR-deficient XPC-negative cells to ensure that, within locally UV-exposed cell nuclei, repair replication was exclusively due to ongoing TCR. In combination with γ-H2AX immunofluorescence labeling (17,24,25) for precise localization of UV-induced DNA-damaged areas, we quantified repair replication in those areas via incorporation of a fluorophore-coupled deoxynucleoside analog into newly synthesized DNA (22) (Fig. 4A and SI Appendix, Fig.…”

Section: Resultsmentioning

confidence: 99%

ELL, a novel TFIIH partner, is involved in transcription restart after DNA repair

Mourgues

Gautier

Lagarou

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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DNA lesions that block transcription may cause cell death even when repaired, if transcription does not restart to reestablish cellular metabolism. However, transcription resumption after individual DNA-lesion repair remains poorly described in mechanistic terms and its players are largely unknown. The general transcription factor II H (TFIIH) is a major actor of both nucleotide excision repair subpathways of which transcription-coupled repair highlights the interplay between DNA repair and transcription. Using an unbiased proteomic approach, we have identified the protein eleven-nineteen lysine-rich leukemia (ELL) as a TFIIH partner. Here we show that ELL is recruited to UV-damaged chromatin in a Cdk7-dependent manner (a component of the cyclin-dependent activating kinase subcomplex of TFIIH). We demonstrate that depletion of ELL strongly hinders RNA polymerase II (RNA Pol II) transcription resumption after lesion removal and DNA gap filling. Lack of ELL was also observed to increase RNA Pol II retention to the chromatin during this process. Identifying ELL as an essential player for RNA Pol II restart during cellular DNA damage response opens the way to obtaining a mechanistic description of transcription resumption after DNA repair.elongation factor | CAK | LEC | Cockayne syndrome | TCR D amage to DNA induced by UV irradiation is repaired by the nucleotide excision repair (NER) system. Three DNA repair-deficient disorders emphasize the importance of NER in genome stability (1). In eukaryotic cells, NER can be divided into two pathways: global genome repair (GGR), repairing lesions throughout the genome, and transcription-coupled repair (TCR), which repairs lesions on the transcribed strand of active genes (2). After damage detection, the basal transcription/repair factor II H (TFIIH), containing the XPB and XPD ATPase/helicases is needed to locally unwind the DNA double helix around the lesion. Furthermore, in vitro UV irradiation elicits a change in the composition of TFIIH. Indeed the majority of TFIIH present on UV-damaged chromatin does not contain the ternary cyclindependent activating kinase (CAK) complex (3). More specifically, the CAK complex is found to be only implicated in TCR but not during GGR, where it seems to be released from the remaining TFIIH components, concomitantly with the recruitment of subsequent NER factors (3). To better our understanding of TFIIH functions in vivo, we have used a combination of improved immunoprecipitation assays and proteomic analysis to identify and characterize TFIIH-interacting partners.Our quantitative proteomic approach has identified several putative TFIIH partners, of which the four most enriched constitute the Little Elongation Complex (LEC): eleven-nineteen lysinerich leukemia (ELL), EAF1 (ELL-associated factor 1), KIAA0947, and NARG2 (4). The proposed role of the LEC is to regulate the transcription of small nuclear RNA genes (5). Although ELL and EAF1 are also part of the Super Elongation Complex (SEC), known to regulate the transcriptional elongation c...

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Section: Resultsmentioning

confidence: 99%

ELL, a novel TFIIH partner, is involved in transcription restart after DNA repair

Mourgues

Gautier

Lagarou

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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“…Although ATR activation has been suggested to occur in non-cycling cells (25)(26)(27)(28)(29)(30)(31)(32)(33), its signaling function has not been extensively examined and its bona fide substrates are not known. Thus, to examine the role of ATR kinase activity in DNA damage signaling in non-cycling cells, we made use of the highly selective ATR inhibitor VE-821 (21,53).…”

Section: Atr Contributes To Dna Damage Response Protein Phosphorylatimentioning

confidence: 99%

“…Although previous studies have indicated that ATR can be activated in noncycling cells in response to DNA repair intermediates (25)(26)(27)(28)(29)(30)(31)(32) and in response to the stalling of RNA polymerase (33)(34)(35), the functional significance of ATR in these contexts in non-cycling cells has not been examined.…”

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confidence: 99%

ATR Kinase Inhibition Protects Non-cycling Cells from the Lethal Effects of DNA Damage and Transcription Stress

Kemp

Sancar

2016

Journal of Biological Chemistry

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ATR (ataxia telangiectasia and Rad-3-related) is a protein kinase that maintains genome stability and halts cell cycle phase transitions in response to DNA lesions that block DNA polymerase movement. These DNA replication-associated features of ATR function have led to the emergence of ATR kinase inhibitors as potential adjuvants for DNA-damaging cancer chemotherapeutics. However, whether ATR affects the genotoxic stress response in non-replicating, non-cycling cells is currently unknown. We therefore used chemical inhibition of ATR kinase activity to examine the role of ATR in quiescent human cells. Although ATR inhibition had no obvious effects on the viability of non-cycling cells, inhibition of ATR partially protected nonreplicating cells from the lethal effects of UV and UV mimetics. Analyses of various DNA damage response signaling pathways demonstrated that ATR inhibition reduced the activation of apoptotic signaling by these agents in non-cycling cells. The pro-apoptosis/cell death function of ATR is likely due to transcription stress because the lethal effects of compounds that block RNA polymerase movement were reduced in the presence of an ATR inhibitor. These results therefore suggest that whereas DNA polymerase stalling at DNA lesions activates ATR to protect cell viability and prevent apoptosis, the stalling of RNA polymerases instead activates ATR to induce an apoptotic form of cell death in non-cycling cells. These results have important implications regarding the use of ATR inhibitors in cancer chemotherapy regimens.Ultraviolet (UV) photoproducts and other bulky DNA lesions block the progression of DNA and RNA polymerases and lead to the activation of various DNA damage responses that are regulated by the action of the ataxia telangiectasia and Rad-3-related (ATR) 3 protein kinase. For example, in response to DNA polymerase stalling and uncoupling from DNA helicase activity (1), ATR is thought to be recruited to singlestranded regions of DNA where it promotes a number of DNA metabolic processes that maintain genomic integrity (2-6), including replication fork stabilization, DNA synthesis and lesion bypass, homologous recombination, replication origin firing, and cell cycle delay. The importance of ATR kinase activity in these responses in replicating cells is evidenced by the fate of cells in which the catalytic activity of ATR has been genetically or pharmacologically inhibited following replication stress, which include apoptosis and entry into catastrophic mitoses (7-10). Thus, ATR helps replicating cells cope with DNA damage and replication stress by facilitating cellular processes that maintain genomic stability and avoid cell death. Given these features of ATR, there has been great interest in the development and use of selective small-molecule inhibitors of ATR kinase activity in cancer chemotherapy regimens in conjunction with compounds that damage DNA and generate replication stress (11-13). For example, several studies have shown that ATR inhibition sensitizes cancer cells to cell kill...

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“…For example, stalled RNA polymerase II may stimulate p53-mediated apoptosis via ATR, even in the absence of replication [148][149][150], possibly via accumulation of RPA on regions of single-stranded DNA [151]. Activation of PARP at SSBs can trigger another mechanism of cell death.…”

Section: Single-strand Lesions and Neuronal Cell Deathmentioning

confidence: 99%

DNA strand break repair and neurodegeneration

Rulten

Caldecott

2013

DNA Repair

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A number of DNA repair disorders are known to cause neurological problems. These disorders can be broadly characterised into early developmental, mid-to-late developmental or progressive. The exact developmental processes that are affected can influence disease pathology, with symptoms ranging from early embryonic lethality to late-onset ataxia. The category these diseases belong to depends on the frequency of lesions arising in the brain, the role of the defective repair pathway, and the nature of the mutation within the patient. Using observations from patients and transgenic mice, we discuss the importance of double strand break repair during neuroprogenitor proliferation and brain development and the repair of single stranded lesions in neuronal function and maintenance.

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UV-induced photolesions elicit ATR-kinase-dependent signaling in non-cycling cells through nucleotide excision repair-dependent and -independent pathways

Cited by 95 publications

References 64 publications

ELL, a novel TFIIH partner, is involved in transcription restart after DNA repair

ELL, a novel TFIIH partner, is involved in transcription restart after DNA repair

ATR Kinase Inhibition Protects Non-cycling Cells from the Lethal Effects of DNA Damage and Transcription Stress

DNA strand break repair and neurodegeneration

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