Interplay between Np95 and Eme1 in the DNA damage response

Mistry, Helena; Gibson, Lianne; Yun, Ji Weon; Sarras, Haya; Tamblyn, Laura; McPherson, J. Peter

doi:10.1016/j.bbrc.2008.07.146

Cited by 18 publications

(15 citation statements)

References 28 publications

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“…As shown in Fig. 1A, we found that the UHRF1 protein level is also regulated in response to DNA damage, i.e., the UHRF1 protein level is reduced upon UV irradiation, consistent with a proposed role of UHRF1 in the DDR (16,20,(25)(26)(27). The reduction of UHRF1 in the DDR is likely attributable to proteasome-mediated degradation, because the proteasome inhibitor MG132 restored Interestingly, the USP7 level and its interaction with UHRF1 remained unaltered in the DDR (Fig.…”

Section: Resultssupporting

confidence: 75%

DNA Damage Regulates UHRF1 Stability via the SCF^β-TrCP E3 Ligase

Chen

Inuzuka

et al. 2013

Molecular and Cellular Biology

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f UHRF1 (ubiquitin-like, with PHD and RING finger domains 1) is a critical epigenetic player involved in the maintenance of DNA methylation patterns during DNA replication. Dysregulation of the UHRF1 level is implicated in cancer onset, metastasis, and tumor recurrence. Previous studies demonstrated that UHRF1 can be stabilized through USP7-mediated deubiquitylation, but the mechanism through which UHRF1 is ubiquitylated is still unknown. Here we show that proteasomal degradation of UHRF1 is mediated by the SCF ␤-TrCP E3 ligase. Through bioinformatic and mutagenesis studies, we identified a functional DSG degron in the UHRF1 N terminus that is necessary for UHRF1 stability regulation. We further show that UHRF1 physically interacts with ␤-TrCP1 in a manner dependent on phosphorylation of serine 108 (S108 UHRF1 ) within the DSG degron. Furthermore, we demonstrate that S108 UHRF1 phosphorylation is catalyzed by casein kinase 1 delta (CK1␦) and is important for the recognition of UHRF1 by SCF ␤-TrCP . Importantly, we demonstrate that UHRF1 degradation is accelerated in response to DNA damage, coincident with enhanced S108 UHRF1 phosphorylation. Taken together, our data identify SCF ␤-TrCP as a bona fide UHRF1 E3 ligase important for regulating UHRF1 steady-state levels both under normal conditions and in response to DNA damage.T he epigenetic regulator UHRF1 is composed of multiple functional domains, including the UBL, Tudor, PHD, SRA, and RING domains, which are responsible for the recognition of histone and DNA methylation as well as ubiquitylation by UHRF1. These domains underlie the ability of UHRF1 to play a role in multiple processes, such as maintenance of DNA methylation, heterochromatin organization, and gene transcription (1-8). Previous studies identified a correlation between UHRF1 overexpression and cancer progression and metastasis, possibly through silencing of various tumor suppressor genes (9-12). Moreover, UHRF1 is implicated in apoptosis in response to DNA damage. Murine embryonic stem cells with targeted disruption of the Uhrf1 gene are hypersensitive to DNA-damaging agents (13). Similarly, knockdown of UHRF1 in HEK293 and WI-38 cells also renders these cells hypersensitive to X rays, UV light, and hydroxyurea (14). More recently, UHRF1 has also been shown to facilitate the DNA damage response (DDR) to gamma irradiation (15, 16). Consistently, DNA damage results in a decrease in the UHRF1 mRNA as well as protein level (1). More recent studies suggest that UHRF1 turnover is controlled by proteasome-mediated degradation. These studies identified the deubiquitylase USP7 in the regulation of the UHRF1 level in vivo (17-19). Specifically, UHRF1 is protected from proteasome-mediated degradation through its association with the deubiquitylase USP7, in a cell cycle-dependent manner. At the M phase of the cell cycle, USP7 disassociates from UHRF1, thus exposing UHRF1 to proteasomal degradation (18). Importantly, manipulating the UHRF1 level in cells has been shown to affect cell proliferation (11,18,20...

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Section: Resultssupporting

confidence: 75%

DNA Damage Regulates UHRF1 Stability via the SCF^β-TrCP E3 Ligase

Chen

Inuzuka

et al. 2013

Molecular and Cellular Biology

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“…Accordingly, we examined UHRF1 protein-protein interactions and found both ERCC1/XPF and MUS81/EME1 are associated with UHRF1, suggesting that structure-specific endonucleases interact with UHRF1 in their heterodimeric forms. A protein-protein interaction between UHRF1 and EME1 was previously detected by a yeast two-hybrid screen (Mistry et al, 2008). This observation also suggests a direct interaction between UHRF1 and lesion processing enzymes.…”

Section: Discussionmentioning

confidence: 99%

UHRF1 Contributes to DNA Damage Repair as a Lesion Recognition Factor and Nuclease Scaffold

et al. 2015

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Summary We identified UHRF1 as a binding factor for DNA interstrand crosslink (ICL) lesions through affinity purification of ICL-recognition activities. UHRF1 is recruited to DNA lesions in vivo and binds directly to ICL-containing DNA. UHRF1-deficient cells display increased sensitivity to a variety of DNA damages. We found that loss of UHRF1 led to retarded lesion processing and reduced recruitment of ICL repair nucleases to the site of DNA damage. UHRF1 interacts physically with both ERCC1 and MUS81, two nucleases involved in the repair of ICL lesions. Depletion of both UHRF1 and components of the Fanconi anemia pathway resulted in increased DNA damage sensitivity compared to defect of each mechanism alone. These results suggest that UHRF1 promotes recruitment of lesion-processing activities via its DNA damage recognition affinity and functions as a nuclease recruitment scaffold in parallel to the Fanconi anemia pathway.

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“…In one study, repair factors including PARP-1, XRCC1, TopBP1, PCNA were found in complex with UHRF1 [4]. Additionally, Eme1 a component of an endonuclease required for damage repair is reported to associate with UHRF1 [37]. Therefore, loss of UHRF1 may lead to a defective replication product by any of these mechanisms, activating the damage response pathway and arresting the cell cycle in G2/M (Figure 8).…”

Section: Discussionmentioning

confidence: 99%

UHRF1 depletion causes a G2/M arrest, activation of DNA damage response and apoptosis

Tien

SenBanerjee

Kulkarni

et al. 2011

Biochemical Journal

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SYNOPSIS Ubiquitin-like protein, containing PHD and RING finger domains-1 (UHRF1) is required for cell cycle progression and epigenetic regulation. In this study, we show that depleting cancer cells of UHRF1 causes activation of the DNA damage response pathway, cell cycle arrest in G2/M and apoptosis dependent on caspase-8. The DNA damage response in cells depleted of UHRF1 is illustrated by: phosphorylation of histone H2AX on serine 139, phosphorylation of CHK2 on threonine 68, phosphorylation of CDC25 on serine 216 and phosphorylation of CDK1 on tyrosine 15. Moreover, we find that UHRF1 accumulates at sites of DNA damage suggesting that the cell cycle block in UHRF1 depleted cells is due to an important role in damage repair. The consequence of UHRF1 depletion is apoptosis: cells undergo activation of caspases 8 and 3 and depletion of caspase-8 prevents cell death induced by UHRF1 knock-down. Interestingly, the cell cycle block and apoptosis occurs in p53 containing and deficient cells. From these studies we conclude that UHRF1 links epigenetic regulation with DNA replication.

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Interplay between Np95 and Eme1 in the DNA damage response

Cited by 18 publications

References 28 publications

DNA Damage Regulates UHRF1 Stability via the SCF^β-TrCP E3 Ligase

DNA Damage Regulates UHRF1 Stability via the SCF^β-TrCP E3 Ligase

UHRF1 Contributes to DNA Damage Repair as a Lesion Recognition Factor and Nuclease Scaffold

UHRF1 depletion causes a G2/M arrest, activation of DNA damage response and apoptosis

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Interplay between Np95 and Eme1 in the DNA damage response

Cited by 18 publications

References 28 publications

DNA Damage Regulates UHRF1 Stability via the SCFβ-TrCP E3 Ligase

DNA Damage Regulates UHRF1 Stability via the SCFβ-TrCP E3 Ligase

UHRF1 Contributes to DNA Damage Repair as a Lesion Recognition Factor and Nuclease Scaffold

UHRF1 depletion causes a G2/M arrest, activation of DNA damage response and apoptosis

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DNA Damage Regulates UHRF1 Stability via the SCF^β-TrCP E3 Ligase

DNA Damage Regulates UHRF1 Stability via the SCF^β-TrCP E3 Ligase