Human DNA polymerase η activity and translocation is regulated by phosphorylation

Chen, Yih Wen; Cleaver, James E.; Hatahet, Zafer; Honkanen, Richard E.; Chang, Jang Yang; Yen, Yun; Chou, Kai Ming

doi:10.1073/pnas.0808589105

Cited by 61 publications

(56 citation statements)

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“…Embryos from WT and pol η −/− mice were harvested on day 13.5 for the generation of MEF cells (54). The established pol η −/− MEF cells were transfected with either WT human pol η (hPol η WT) or the R61F mutant form of human pol η (hPol η R61F) (55). The protein expression level of pol η was examined by Western blot analysis with anti-human or anti-mouse pol η antibodies from Biorbyt.…”

Section: Methodsmentioning

confidence: 99%

Ablation of XP-V gene causes adipose tissue senescence and metabolic abnormalities

Chen

Harris

Hatahet

et al. 2015

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

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Obesity and the metabolic syndrome have evolved to be major health issues throughout the world. Whether loss of genome integrity contributes to this epidemic is an open question. DNA polymerase η (pol η), encoded by the xeroderma pigmentosum (XP-V) gene, plays an essential role in preventing cutaneous cancer caused by UV radiation-induced DNA damage. Herein, we demonstrate that pol η deficiency in mice (pol η −/− ) causes obesity with visceral fat accumulation, hepatic steatosis, hyperleptinemia, hyperinsulinemia, and glucose intolerance. In comparison to WT mice, adipose tissue from pol η −/− mice exhibits increased DNA damage and a greater DNA damage response, indicated by upregulation and/or phosphorylation of ataxia telangiectasia mutated (ATM), phosphorylated H 2 AX (γH 2 AX), and poly[ADP-ribose] polymerase 1 (PARP-1). Concomitantly, increased cellular senescence in the adipose tissue from pol η −/− mice was observed and measured by up-regulation of senescence markers, including p53, p16 Ink4a , p21, senescence-associated (SA) β-gal activity, and SA secretion of proinflammatory cytokines interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) as early as 4 wk of age. Treatment of pol η −/− mice with a p53 inhibitor, pifithrin-α, reduced adipocyte senescence and attenuated the metabolic abnormalities. Furthermore, elevation of adipocyte DNA damage with a high-fat diet or sodium arsenite exacerbated adipocyte senescence and metabolic abnormalities in pol η −/− mice. In contrast, reduction of adipose DNA damage with N-acetylcysteine or metformin ameliorated cellular senescence and metabolic abnormalities. These studies indicate that elevated DNA damage is a root cause of adipocyte senescence, which plays a determining role in the development of obesity and insulin resistance.T he human genome is constantly challenged by exogenous and endogenous DNA damaging agents. To ensure genome integrity, human cells have faithful DNA replication machinery and DNA repair systems that are coordinated by DNA damage response networks. In response to different extents or types of DNA lesions, the DNA damage response activates appropriate cellular responses, including transient or permanent (senescence) cell cycle arrest, or apoptosis, to minimize the detrimental effects of DNA lesions (1). Reduction or deficiency in DNA repair/replication enzyme activity is well documented to increase vulnerability for the development of cancer, neurodegenerative diseases, and aging (2). In addition, defective DNA repair enzymes are associated with the metabolic symptom; for example, DNA glycosylase (Neil1)-and OGG1-deficient mice are obese (3-5), and nucleotide excision repair protein ERCC1-XPF deficiency causes lipodystrophy (6). Furthermore, DNA damage response protein ataxia telangiectasia mutated (ATM) suppresses JNK activity through p53 signaling and mediates an antioxidant action that has been suggested to be relevant to the metabolic syndrome (7). Nucleotide excision repair XP-A protein may affect metabolism by altering mitochondrial...

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Section: Methodsmentioning

confidence: 99%

Ablation of XP-V gene causes adipose tissue senescence and metabolic abnormalities

Chen

Harris

Hatahet

et al. 2015

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

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“…After UV irradiation, the monoubiquitination of pol is down-regulated to expose the PCNA-interacting surface, thereby stimulating pol 's interaction with PCNA and facilitating TLS (17). In addition, previous studies showed that pol can be phosphorylated at serine 587, threonine 617, and serine 601 (Ser 601 ), which are required for normal survival and efficient recovery from UV damage (18,19). It is therefore important to identify other possible posttranslational modification(s) of hu-man pol for further understanding the regulatory mechanisms of this important TLS polymerase.…”

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confidence: 99%

The Functions of Serine 687 Phosphorylation of Human DNA Polymerase η in UV Damage Tolerance

Dai

You

Wang

2016

Molecular & Cellular Proteomics

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DNA polymerase (pol ) is a Y-family translesion synthesis polymerase that plays a key role in the cellular tolerance toward UV irradiation-induced DNA damage. Here, we identified, for the first time, the phosphorylation of serine 687 (Ser 687 ), which is located in the highly conserved nuclear localization signal (NLS) region of human pol and is mediated by cyclin-dependent kinase 2 (CDK2). We also showed that this phosphorylation is stimulated in human cells upon UV light exposure and results in diminished interaction of pol with proliferating cell nuclear antigen (PCNA). Furthermore, we demonstrated that the phosphorylation of Ser 687 in pol confers cellular protection from UV irradiation and increases the efficiency in replication across a site-specifically incorporated cyclobutane pyrimidine dimer in human cells. Based on these results, we proposed a mechanistic model where Ser 687 phosphorylation functions in the reverse polymerase switching step of translesion synthesis: The phosphorylation brings negative charges to the NLS of pol , which facilitates its departure from PCNA, thereby resetting the replication fork for highly accurate and processive DNA replication. Thus, our study, together with previous findings, supported that the posttranslational modifications of NLS of pol played a dual role in polymerase switching, where Lys 682 deubiquitination promotes the recruitment of pol to PCNA immediately prior to lesion bypass and Ser 687 phosphorylation stimulates its departure from the replication fork immediately after lesion bypass. Molecular & Cellular

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“…Platinum compounds react with DNA and lead to DNA lesions, including intrastrand crosslinks (Pt-d[CpG], Pt-d [ApG] and Pt-d [GpNgG]), interstrand crosslinks and single nucleotide damage involving guanine [43] . Hence, TLS that allows bypass of the intrastrand crosslinks and constitutes a critical initial step in interstrand crosslink repair [44] is advantageous to tumor cells' survival [45][46][47][48] . These crosslink bypasses can result in resistance against platinum-based chemotherapy [47,49] .…”

Section: Discussionmentioning

confidence: 99%

RAD18 polymorphisms are associated with platinum-based chemotherapy toxicity in Chinese patients with non-small cell lung cancer

Chu

Shao

et al. 2016

Acta Pharmacol Sin

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Aim: Although targeted therapy is very efficient for lung cancer, traditional platinum-based chemotherapies are still the principal strategy in the absence of positive biomarkers. The aim of the present study is to evaluate the contribution of RAD18 polymorphisms to platinum-chemotherapy response and its potential side effects in Chinese patients with non-small cell lung cancer (NSCLC). Methods: A total of 1021 Chinese patients with histological diagnosis of advanced NSCLC were enrolled. Treatment responses were classified into 4 categories (complete response, partial response, stable disease and progressive disease). Gastrointestinal and hematological toxicity incidences were assessed twice a week during the first-line treatment. Ten RAD18 SNPs were genotyped. A logistic regression model was utilized to analyze the associations between RAD18 SNPs and treatment response or toxicity. Results: Among the 10 SNPs tested, none was significantly correlated with the treatment response in a combined cohort. For gastrointestinal toxicity incidences, rs586014 was significantly associated with an increased risk of grade 3 or 4 gastrointestinal toxicity in non-smokers and in the combined cohort; rs654448 and rs618784 were significantly associated with gastrointestinal toxicity in non-smokers; rs6763823 was significantly associated with gastrointestinal toxicity in smokers. For hematological toxicity incidences, rs586014, rs654448 and rs618784 were significantly associated with hematologic toxicity in non-smokers; rs6763823 and rs9880051 were significantly associated with leukocytopenia in smokers. Conclusion: RAD18 polymorphisms are correlated with the side effects of platinum-chemotherapy in Chinese patients with advanced NSCLC.

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Human DNA polymerase η activity and translocation is regulated by phosphorylation

Cited by 61 publications

References 50 publications

Ablation of XP-V gene causes adipose tissue senescence and metabolic abnormalities

Ablation of XP-V gene causes adipose tissue senescence and metabolic abnormalities

The Functions of Serine 687 Phosphorylation of Human DNA Polymerase η in UV Damage Tolerance

RAD18 polymorphisms are associated with platinum-based chemotherapy toxicity in Chinese patients with non-small cell lung cancer

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