234 results on '"Exo1"'
Search Results
2. Novel candidate metastasis‐associated genes for synovial sarcoma.
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Zhao, Zhiqing, Niu, Jianfang, Wang, Jichuan, Zhang, Ranxin, Liang, Haijie, Ma, Yingteng, Ferrena, Alexander, Wang, Wei, Yang, Rui, Geller, David S., Guo, Wei, Ren, Tingting, Hoang, Bang H., Tang, Xiaodong, and Yan, Taiqiang
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HOMOLOGOUS recombination ,SYNOVIOMA ,GENE expression ,SARCOMA ,GENE regulatory networks - Abstract
Synovial sarcoma (SS) is an aggressive soft tissue sarcoma with poor prognosis due to late recurrence and metastasis. Metastasis is an important prognostic factor of SS. This study aimed to identify the core genes and mechanisms associated with SS metastasis. Microarray data for GSE40021 and GSE40018 were obtained from the Gene Expression Omnibus database. 186 differentially expressed genes (DEGs) were identified. The biological functions and signalling pathways closely associated with SS metastasis included extracellular matrix (ECM) organization and ECM‐receptor interaction. Gene set enrichment analysis showed that the terms cell cycle, DNA replication, homologous recombination and mismatch repair were significantly enriched in the metastasis group. Weighted gene co‐expression network analysis identified the most relevant module and 133 hub genes, and 31 crossover genes were identified by combining DEGs. Subsequently, four characteristic genes, EXO1, NCAPG, POLQ and UHRF1, were identified as potential biomarkers associated with SS metastasis using the least absolute shrinkage and selection operator algorithm and validation dataset verification analysis. Immunohistochemistry results from our cohort of 49 patients revealed visible differences in the expression of characteristic genes between the non‐metastatic and metastatic groups. Survival analysis indicated that high expression of characteristic genes predicted poor prognosis. Our data revealed that primary SS samples from patients who developed metastasis showed activated homologous recombination and mismatch repair compared to samples from patients without metastasis. Furthermore, EXO1, NCAPG, POLQ and UHRF1 were identified as potential candidate metastasis‐associated genes. This study provides further research insights and helps explore the mechanisms of SS metastasis. [ABSTRACT FROM AUTHOR]
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- 2024
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3. PARG-deficient tumor cells have an increased dependence on EXO1/FEN1-mediated DNA repair.
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Andronikou, Christina, Burdova, Kamila, Dibitetto, Diego, Lieftink, Cor, Malzer, Elke, Kuiken, Hendrik J, Gogola, Ewa, Ray Chaudhuri, Arnab, Beijersbergen, Roderick L, Hanzlikova, Hana, Jonkers, Jos, and Rottenberg, Sven
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SINGLE-strand DNA breaks , *DNA repair , *DNA mismatch repair , *CD38 antigen , *DNA replication , *BRCA genes - Abstract
Targeting poly(ADP-ribose) glycohydrolase (PARG) is currently explored as a therapeutic approach to treat various cancer types, but we have a poor understanding of the specific genetic vulnerabilities that would make cancer cells susceptible to such a tailored therapy. Moreover, the identification of such vulnerabilities is of interest for targeting BRCA2;p53-deficient tumors that have acquired resistance to poly(ADP-ribose) polymerase inhibitors (PARPi) through loss of PARG expression. Here, by performing whole-genome CRISPR/Cas9 drop-out screens, we identify various genes involved in DNA repair to be essential for the survival of PARG;BRCA2;p53-deficient cells. In particular, our findings reveal EXO1 and FEN1 as major synthetic lethal interactors of PARG loss. We provide evidence for compromised replication fork progression, DNA single-strand break repair, and Okazaki fragment processing in PARG;BRCA2;p53-deficient cells, alterations that exacerbate the effects of EXO1/FEN1 inhibition and become lethal in this context. Since this sensitivity is dependent on BRCA2 defects, we propose to target EXO1/FEN1 in PARPi-resistant tumors that have lost PARG activity. Moreover, EXO1/FEN1 targeting may be a useful strategy for enhancing the effect of PARG inhibitors in homologous recombination-deficient tumors. Synopsis: Targeting poly(ADP-ribose) glycohydrolase (PARG) is being explored as anti-cancer therapeutic strategy, and PARG loss may also contribute to resistance to PARP inhibitor (PARPi) treatment. This study provides insights into specific vulnerabilities that can render homologous-recombination (HR)-deficient tumors susceptible to the loss of PARG activity. Impaired de-PARylation in PARG-deficient cells affects the repair of single-strand breaks (SSBs) and processing of unligated Okazaki fragments. Genome-wide CRISPR/Cas9 screens reveal EXO1 and FEN1 as critical factors in cells deficient for PARG and BRCA2. Inhibition of EXO1/FEN1 in PARG;BRCA2-deficient cells results in unresolved Okazaki fragments, which persist as single-strand DNA (ssDNA) gaps. Persistent ssDNA gaps become specifically lethal to HR-deficient cells when converted into double-strand breaks (DSBs) upon replication. Genome-wide loss-of-function screens reveal DNA repair genes essential in HR-defective cells lacking PARG, suggesting ways for therapeutically exploiting PARG inhibitors and targeting PARPi-resistant tumors. [ABSTRACT FROM AUTHOR]
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- 2024
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4. EXO1/P53/SREBP1 axis-regulated lipid metabolism promotes prostate cancer progression
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Zefeng Wang, Zheng Chao, Qi Wang, Fan Zou, Tianbao Song, Lizhe Xu, Jinzhuo Ning, and Fan Cheng
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Prostate cancer ,Lipid metabolism ,EXO1 ,P53 ,Biomarker ,Medicine - Abstract
Abstract Prostate cancer (PCa) is one of the most common malignant tumors affecting the male genitourinary system. However, there is currently a lack of effective treatments for patients with advanced prostate cancer, which significantly impacts men’s overall health. Exonuclease 1 (EXO1), a protein with mismatch repair and recombination functions, has been found to play a vital role in various diseases. In our study, we discovered that EXO1 acts as a novel biomarker of PCa, which promotes prostate cancer progression by regulating lipid metabolism reprogramming in prostate cancer cells. Mechanistically, EXO1 promotes the expression of SREBP1 by inhibiting the P53 signaling pathway. In summary, our findings suggest that EXO1 regulated intracellular lipid reprogramming through the P53/SREBP1 axis, thus promoting PCa progression. The result could potentially lead to new insights and therapeutic targets for diagnosing and treating PCa.
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- 2024
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5. EXO1/P53/SREBP1 axis-regulated lipid metabolism promotes prostate cancer progression.
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Wang, Zefeng, Chao, Zheng, Wang, Qi, Zou, Fan, Song, Tianbao, Xu, Lizhe, Ning, Jinzhuo, and Cheng, Fan
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PROSTATE cancer , *LIPID metabolism , *CANCER invasiveness , *PROSTATE cancer patients , *METABOLIC reprogramming , *CANCER patients - Abstract
Prostate cancer (PCa) is one of the most common malignant tumors affecting the male genitourinary system. However, there is currently a lack of effective treatments for patients with advanced prostate cancer, which significantly impacts men's overall health. Exonuclease 1 (EXO1), a protein with mismatch repair and recombination functions, has been found to play a vital role in various diseases. In our study, we discovered that EXO1 acts as a novel biomarker of PCa, which promotes prostate cancer progression by regulating lipid metabolism reprogramming in prostate cancer cells. Mechanistically, EXO1 promotes the expression of SREBP1 by inhibiting the P53 signaling pathway. In summary, our findings suggest that EXO1 regulated intracellular lipid reprogramming through the P53/SREBP1 axis, thus promoting PCa progression. The result could potentially lead to new insights and therapeutic targets for diagnosing and treating PCa. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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6. Investigating the importance of EGFR (− 216G/T), Exo1 (K589E) and LEP (− 2548G/A) gene polymorphisms with risk of lung cancer as potential diagnostic biomarker in Iranian population
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Milad Pezeshki, Sayed Mostafa Hosseini, Jamshid Ansari, and Azam Ahmadi
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Lung cancer ,Early genetically detection ,SNPs ,EGFR ,Exo1 ,LEP ,Medicine (General) ,R5-920 ,Genetics ,QH426-470 - Abstract
Abstract Background In Iran, lung cancer is the third most common type of cancer and its prevalence is increasing rapidly. Identification informative genetic polymorphisms in cancer causing genes including epidermal growth factor receptor (EGFR) as key gene in control of cellular proliferation via intrinsic tyrosine/kinase activity, exonuclease 1 (EXO1) as one of the upregulated gene in different human malignancies and leptin (LEP) participate in carcinogenesis in lung cancer appears to be used as potential genetic markers for predicting lung cancer risk. There is no study about investigate association of the EGFR (− 216G/T), Exo1 (K589E) and LEP (− 2548G/A) gene polymorphisms with risk of lung cancer in Iranian population. The aim of this study was investigating the association of EGFR (− 216G/T), Exo1 (K589E) and LEP (− 2548G/A) gene polymorphisms with risk of lung cancer as a potentially diagnostic biomarker in Iranian population. Methods In this case–control study, A total of 100 patients with lung cancer and 100 age and gender-matched healthy controls were recruited into this study and the association between EGFR (− 216G/T), Exo1 (K589E) and LEP (− 2548G/A) gene polymorphisms with the risk of lung cancer was investigated by using PCR–RFLP technique and bioinformatics approach. Results The rs712829 of EGFR gene show that a significant statistical difference between G allele and risk of lung cancer (P = 0.001, OR = 2.976, CI = 95%, 1.557–5.691), in contrast, the T allele and TT genotype show a protective role against the risk of lung cancer. Result of in silico analysis indicated that the rs712829 alter splicing and promoter regulation of EGFR gene and associated with the risk of lung cancer. PCR–RFLP result for rs1047840 of Exo1 gene showed that the AA genotype and A allele of this polymorphism associated with risk of lung cancer, whereas the GG genotype show a protective effect against the risk of lung cancer (P = 0.004, OR = 5.391, CI = 95%, 1.690–17.200). On the other hand, in silico analysis showed that the existence of rs1047840 in Exo1 gene influence lung cancer susceptibility. For rs7799039 of LEP gene, PCR–RFLP analysis showed that, there is no significant association between this polymorphism and the risk of lung cancer. Conclusion The rs712829 of EGFR gene and rs1047840 of Exo1 are associated with risk of lung cancer among Iranian population and can be used as a potentially candidate biomarker for early detection and primary prevention.
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- 2023
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7. The FBXO32/ATR/ATM axis acts as a molecular switch to control the sensitivity of osteosarcoma cells to irradiation through its regulation of EXO1 expression
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Lu Yao, Huang Panpan, Li Yanli, Liu Wenyu, Li Jing, Zhao Rui, Feng Haihua, Shi Ce, and Cao Gaolu
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osteosarcoma ,EXO1 ,ATM ,ATR ,FBXO32 ,irradiation ,Biochemistry ,QD415-436 ,Genetics ,QH426-470 - Abstract
Osteosarcoma (OS) is the most common primary bone cancer in children and adolescents. In clinical treatments, the insensitivity of OS to conventional radiotherapy regimens significantly contributes to poor patient prognosis and survival. EXO1 is responsible for DNA repair pathways and telomere maintenance. Meanwhile, ATM and ATR are considered switches because they can regulate the expression of EXO1. However, their expression and interaction in OS cells under irradiation (IR) remain unclear. This study aims to investigate the roles of FBXO32, ATM, ATR and EXO1 in OS radiotherapy insensitivity and poor patient prognosis and explore potential pathogenic mechanisms. Bioinformatics is employed to analyse differential gene expression and correlations with prognosis in OS. Cell counting kit 8 assay, clone formation assay, and flow cytometry are used to evaluate cell survival and apopotosis under IR. Co-IP assay is used to detect protein‒protein interactions. Bioinformatics analysis reveals that EXO1 is closely related to survival, apoptosis and poor prognosis in OS. Silencing of EXO1 suppresses cell proliferation and increases the sensitivity of OS cells. Molecular biological experiments show that ATM and ATR act as switches to regulate EXO1 expression under IR. Higher expression of EXO1, which is closely correlated with IR insensitivity and poorer prognosis, might be used as a prognostic indicator for OS. Phosphorylated ATM enhances the expression of EXO1, and phosphorylated ATR induces the degradation of EXO1. More importantly, FBXO32 degrades ATR via ubiquitination in a time-dependent manner. Our data may provide a reference for future research in the mechanisms, clinical diagnosis, and treatment of OS.
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- 2023
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8. Investigating the importance of EGFR (− 216G/T), Exo1 (K589E) and LEP (− 2548G/A) gene polymorphisms with risk of lung cancer as potential diagnostic biomarker in Iranian population.
- Author
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Pezeshki, Milad, Hosseini, Sayed Mostafa, Ansari, Jamshid, and Ahmadi, Azam
- Subjects
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GENETIC polymorphisms , *LEPTIN receptors , *IRANIANS , *LUNG cancer , *DISEASE risk factors , *EPIDERMAL growth factor receptors , *CANCER genes - Abstract
Background: In Iran, lung cancer is the third most common type of cancer and its prevalence is increasing rapidly. Identification informative genetic polymorphisms in cancer causing genes including epidermal growth factor receptor (EGFR) as key gene in control of cellular proliferation via intrinsic tyrosine/kinase activity, exonuclease 1 (EXO1) as one of the upregulated gene in different human malignancies and leptin (LEP) participate in carcinogenesis in lung cancer appears to be used as potential genetic markers for predicting lung cancer risk. There is no study about investigate association of the EGFR (− 216G/T), Exo1 (K589E) and LEP (− 2548G/A) gene polymorphisms with risk of lung cancer in Iranian population. The aim of this study was investigating the association of EGFR (− 216G/T), Exo1 (K589E) and LEP (− 2548G/A) gene polymorphisms with risk of lung cancer as a potentially diagnostic biomarker in Iranian population. Methods: In this case–control study, A total of 100 patients with lung cancer and 100 age and gender-matched healthy controls were recruited into this study and the association between EGFR (− 216G/T), Exo1 (K589E) and LEP (− 2548G/A) gene polymorphisms with the risk of lung cancer was investigated by using PCR–RFLP technique and bioinformatics approach. Results: The rs712829 of EGFR gene show that a significant statistical difference between G allele and risk of lung cancer (P = 0.001, OR = 2.976, CI = 95%, 1.557–5.691), in contrast, the T allele and TT genotype show a protective role against the risk of lung cancer. Result of in silico analysis indicated that the rs712829 alter splicing and promoter regulation of EGFR gene and associated with the risk of lung cancer. PCR–RFLP result for rs1047840 of Exo1 gene showed that the AA genotype and A allele of this polymorphism associated with risk of lung cancer, whereas the GG genotype show a protective effect against the risk of lung cancer (P = 0.004, OR = 5.391, CI = 95%, 1.690–17.200). On the other hand, in silico analysis showed that the existence of rs1047840 in Exo1 gene influence lung cancer susceptibility. For rs7799039 of LEP gene, PCR–RFLP analysis showed that, there is no significant association between this polymorphism and the risk of lung cancer. Conclusion: The rs712829 of EGFR gene and rs1047840 of Exo1 are associated with risk of lung cancer among Iranian population and can be used as a potentially candidate biomarker for early detection and primary prevention. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
9. LncRNA CECR7 boosts hepatocellular carcinoma progression by recruiting RNA binding protein U2AF2 to enhance the stability of EXO1 mRNA
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Liang Zhao, Qing Zang, Guodong Liang, and Xiaobin Yao
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Hepatocellular carcinoma ,LncRNA ,CECR7 ,EXO1 ,U2AF2 ,Science (General) ,Q1-390 ,Social sciences (General) ,H1-99 - Abstract
Objective: As an important factor tumor regulator,long non-coding RNAs (lncRNAs) have aroused extensive attention via the diverse functional mechanisms that were associated with the pathological and physiological processes of HCC. Here, the main purpose of this study was to provide a clear understanding about the expression, functions and potential mechanism of lncRNA CECR7 (Cat Eye Syndrome Chromosome Region, Candidate 7) in HCC. Methods: RT-qPCR analysis and TCGA database analysis were applied to investigate the expression of CECR7 in HCC cell lines and tissues. Chi-squared Test was employed to explore the correlation between CECR7 expression and HCC clinicopathological features. Besides, Kaplan-Meier curves were constructed to test the effects of CECR7 expression on the prognosis of HCC patients. Transwell assays, MTT assay EdU assay and animal experiments were applied to explore the effects of CECR7 expression on HCC cells migration, invasion, and growth. Furthermore, RNA-seq analysis, luciferase reporter assay and mRNA decay rates assessment were utilized to investigate the mechanism whereby CECR7 regulated EXO1 mRNA. And, rescue experiments were used to determine whether EXO1 was an essential mediator for CECR7 to accelerate HCC cells migration, invasion, and growth. Results: CECR7 was determined to be significantly overexpressed in HCC cell lines and tissues. CECR7 expression was closely correlated with the tumor size, venous infiltration, TNM stage, 5-year overall survival and disease-free survival of HCC. And, CECR7 played a catalytic role in HCC cells migration, invasion, and growth. Furthermore, CECR7 enhanced the stability of EXO1 mRNA by recruiting RNA binding protein U2AF2. And, EXO1 was determined to be an essential mediator for CECR7 to accelerate HCC cells migration, invasion, and growth. Conclusion: In a word, our findings demonstrates that the cancer-promoting gene lncRNA CECR7 motivates HCC metastasis and growth through enhanced mRNA stability of EXO1 mediated by U2AF2, proposing a new insight for targeted therapy of HCC.
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- 2023
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10. Long-range DNA end resection supports homologous recombination by checkpoint activation rather than extensive homology generation
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Michael T Kimble, Matthew J Johnson, Mattie R Nester, and Lorraine S Symington
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homologous recombination ,DNA repair ,checkpoint ,Exo1 ,Sgs1 ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Homologous recombination (HR), the high-fidelity mechanism for double-strand break (DSB) repair, relies on DNA end resection by nucleolytic degradation of the 5′-terminated ends. However, the role of long-range resection mediated by Exo1 and/or Sgs1-Dna2 in HR is not fully understood. Here, we show that Exo1 and Sgs1 are dispensable for recombination between closely linked repeats, but are required for interchromosomal repeat recombination in Saccharomyces cerevisiae. This context-specific requirement for long-range end resection is connected to its role in activating the DNA damage checkpoint. Consistent with this role, checkpoint mutants also show a defect specifically in interchromosomal recombination. Furthermore, artificial activation of the checkpoint partially restores interchromosomal recombination to exo1∆ sgs1∆ cells. However, cell cycle delay is insufficient to rescue the interchromosomal recombination defect of exo1∆ sgs1∆ cells, suggesting an additional role for the checkpoint. Given that the checkpoint is necessary for DNA damage-induced chromosome mobility, we propose that the importance of the checkpoint, and therefore long-range resection, in interchromosomal recombination is due to a need to increase chromosome mobility to facilitate pairing of distant sites. The need for long-range resection is circumvented when the DSB and its repair template are in close proximity.
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- 2023
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11. Mechanism of homology search expansion during recombinational DNA break repair in Saccharomyces cerevisiae.
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Dumont, Agnès, Mendiboure, Nicolas, Savocco, Jérôme, Anani, Loqmen, Moreau, Pierrick, Thierry, Agnès, Modolo, Laurent, Jost, Daniel, and Piazza, Aurèle
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HOMOLOGOUS recombination , *SINGLE-stranded DNA , *RECOMBINANT DNA , *CHROMOSOMES , *CHROMATIN , *DOUBLE-strand DNA breaks , *DNA repair - Abstract
Homology search is a central step of DNA double-strand break (DSB) repair by homologous recombination (HR). How it operates in cells remains elusive. We developed a Hi-C-based methodology to map single-stranded DNA (ssDNA) contacts genome-wide in S. cerevisiae , which revealed two main homology search phases. Initial search conducted by short Rad51-ssDNA nucleoprotein filaments (NPFs) is confined in cis by cohesin-mediated chromatin loop folding. Progressive growth of stiff NPFs enables exploration of distant genomic sites. Long-range resection drives this transition from local to genome-wide search by increasing the probability of assembling extensive NPFs. DSB end-tethering promotes coordinated search by opposite NPFs. Finally, an autonomous genetic element on chromosome III engages the NPF, which stimulates homology search in its vicinity. This work reveals the mechanism of the progressive expansion of homology search that is orchestrated by chromatin organizers, long-range resection, end-tethering, and specialized genetic elements and that exploits the stiff NPF structure conferred by Rad51 oligomerization. [Display omitted] • Stiffening of ssDNA by Rad51 enables genome-wide homology search • Exo1-mediated long-range resection promotes genome-wide homology search • Rad51 filaments perform homology search in a co-aligned configuration • The recombination enhancer focuses homology search in its vicinity Repair of DNA breaks by homologous recombination entails a needle-in-a-haystack search for an intact molecule in the genome and nucleus. Dumont et al. devised a method to map the search, which revealed its gradual expansion from local to genome wide and enabled studying its regulation. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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12. Immune infiltration landscape on prognosis and therapeutic response and relevant epigenetic and transcriptomic mechanisms in lung adenocarcinoma.
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Liangming Zhang, Biwang Jiang, Zhuxiang Lan, Chaomian Yang, Yien Yao, Jie Lin, and Qiu Wei
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TRANSCRIPTOMES ,IMMUNE checkpoint proteins ,EPIGENETICS ,GENE regulatory networks ,ADENOCARCINOMA ,IPILIMUMAB - Abstract
Objective: Lung adenocarcinoma (LUAD) is the most prevalent lung cancer subtype, but its immune infiltration features are not comprehensively understood. To address the issue, the present study was initiated to describe the immune infiltrations across LUAD from cellular compositional, functional, and mechanism perspectives. Methods: We adopted five LUAD datasets (GSE32863, GSE43458, GSE75037, TCGA-LUAD, and GSE72094). Differentially expressed genes between LUAD and controls were selected for co-expression network analysis. Risky immune cell types were determined for classifying LUAD patients as diverse subtypes, followed by a comparison of antitumor immunity and therapeutic response between subtypes. Then, LUAD- and subtype-related key module genes affected by DNA methylation were determined for quantifying a scoring scheme. EXO1 was chosen for functional analysis via in vitro assays. Results: Two immune cell infiltration-based subtypes (C1 and C2) were established across LUAD, with poorer prognostic outcomes and lower infiltration of immune cell types in C1. Additionally, C1 presented higher responses to immune checkpoint blockade and targeted agents (JNK inhibitor VIII, BI-D1870, RO-3306, etc.). The scoring system (comprising GAPDH, EXO1, FYN, CFTR, and KLF4) possessed higher accuracy in estimating patients' prognostic outcomes. EXO1 upregulation contributed to the growth, migration, and invasion of LUAD cells. In addition, EXO1 facilitated PD-L1 and sPD-L1 expression in LUAD cells. Conclusion: Altogether, our findings offer a comprehensive understanding of the immune infiltration landscape on prognosis and therapeutic response of LUAD as well as unveil potential epigenetic and transcriptomic mechanisms, which might assist personalized treatment. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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13. Breast cancer-associated SNP rs72755295 is a cis-regulatory variation for human EXO1
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Qiang Shi, Xing-Yuan Yao, Hong-Yan Wang, Ya-Jie Li, Xin-Xin Zhang, and Chang Sun
- Subjects
Breast cancer ,EXO1 ,rs72755295 ,rs4149909 ,expression regulation ,Genetics ,QH426-470 - Abstract
Abstract Breast cancer is the most common malignant tumor in women. A previous genome-wide association study reports that rs72755295, a SNP locating at intron of EXO1 (exonuclease 1), is associated with breast cancer. Due to the complete linkage disequilibrium between rs72755295 and rs4149909, a nonsynonymous mutation for EXO1, rs4149909 is supposed to be the causal SNP. Since EXO1 is overexpressed in breast carcinoma samples, we hypothesized that the genetic variations in this locus might confer breast cancer risk by regulating EXO1 expression. To substantiate this, a functional genomics study was performed. The dual luciferase assay indicated that G of rs72755295 presents significantly higher relative enhancer activity than A, thus verifying that this SNP can influence gene expression in breast cell. Through chromosome conformation capture it was disclosed that the enhancer containing rs72755295 can interact with the EXO1 promoter. RNA-seq analysis indicated that EXO1 expression is dependent on the rs72755295 genotype. By chromatin immunoprecipitation, the transcription factor PAX6 (paired box 6) was recognized to bind the region spanning rs72755295. In electrophoretic mobility shift assay, G of rs72755295 displays obviously higher binding affinity with nuclear protein than A. Our results indicated that rs72755295 is a cis-regulatory variation for EXO1 and might confer breast cancer risk besides rs4149909.
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- 2022
- Full Text
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14. Genome Replication Is Associated With Release of Immunogenic DNA Waste.
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Schubert, Nadja, Schumann, Tina, Daum, Elena, Flade, Karolin, Ge, Yan, Hagedorn, Lara, Edelmann, Winfried, Müller, Luise, Schmitz, Marc, Kuut, Gunnar, Hornung, Veit, Behrendt, Rayk, and Roers, Axel
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EXONUCLEASES ,DNA ,DNA replication ,ENDONUCLEASES ,SYSTEMIC lupus erythematosus ,GENOMES ,TYPE I interferons - Abstract
Innate DNA sensors detect foreign and endogenous DNA to induce responses to infection and cellular stress or damage. Inappropriate activation by self-DNA triggers severe autoinflammatory conditions, including Aicardi-Goutières syndrome (AGS) that can be caused by defects of the cytosolic DNase 3'repair exonuclease 1 (TREX1). TREX1 loss-of-function alleles are also associated with systemic lupus erythematosus (SLE). Chronic activation of innate antiviral immunity in TREX1-deficient cells depends on the DNA sensor cGAS, implying that accumulating TREX1 DNA substrates cause the inflammatory pathology. Retrotransposon-derived cDNAs were shown to activate cGAS in TREX1-deficient neuronal cells. We addressed other endogenous sources of cGAS ligands in cells lacking TREX1. We find that induced loss of TREX1 in primary cells induces a rapid IFN response that requires ongoing proliferation. The inflammatory phenotype of Trex1
-/- mice was partially rescued by additional knock out of exonuclease 1, a multifunctional enzyme providing 5' flap endonuclease activity for Okazaki fragment processing and postreplicative ribonucleotide excision repair. Our data imply genome replication as a source of DNA waste with pathogenic potential that is efficiently degraded by TREX1. [ABSTRACT FROM AUTHOR]- Published
- 2022
- Full Text
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15. PHF11 promotes DSB resection, ATR signaling, and HR
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Gong, Yi, Handa, Naofumi, Kowalczykowski, Stephen C, and de Lange, Titia
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Biochemistry and Cell Biology ,Biological Sciences ,Genetics ,Cancer ,Animals ,Cell Line ,DNA Breaks ,Double-Stranded ,DNA Repair ,DNA-Binding Proteins ,Exodeoxyribonucleases ,HEK293 Cells ,Homologous Recombination ,Humans ,Mice ,Protein Transport ,Signal Transduction ,Transcription Factors ,PELF11 ,EXO1 ,RPA ,ATR ,DSB ,resection ,homologous recombination ,PHF11 ,Medical and Health Sciences ,Psychology and Cognitive Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences ,Psychology - Abstract
Resection of double-strand breaks (DSBs) plays a critical role in their detection and appropriate repair. The 3' ssDNA protrusion formed through resection activates the ATR-dependent DNA damage response (DDR) and is required for DSB repair by homologous recombination (HR). Here we report that PHF11 (plant homeodomain finger 11) encodes a previously unknown DDR factor involved in 5' end resection, ATR signaling, and HR. PHF11 was identified based on its association with deprotected telomeres and localized to sites of DNA damage in S phase. Depletion of PHF11 diminished the ATR signaling response to telomere dysfunction and genome-wide DNA damage, reduced end resection at sites of DNA damage, resulted in compromised HR and misrejoining of S-phase DSBs, and increased the sensitivity to DNA-damaging agents. PHF11 interacted with the ssDNA-binding protein RPA and was found in a complex with several nucleases, including the 5' dsDNA exonuclease EXO1. Biochemical experiments demonstrated that PHF11 stimulates EXO1 by overcoming its inhibition by RPA, suggesting that PHF11 acts (in part) by promoting 5' end resection at RPA-bound sites of DNA damage. These findings reveal a role for PHF11 in DSB resection, DNA damage signaling, and DSB repair.
- Published
- 2017
16. Exonuclease 1 is a Potential Diagnostic and Prognostic Biomarker in Hepatocellular Carcinoma
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Jiaxiu Ma, Jiapei Jin, Huishuang Lu, Jin Zhang, Yalan Li, and Xuefei Cai
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Exo1 ,hepatocellular carcinoma ,clinical outcome ,immune cell infiltration ,immune checkpoints ,DNA methylation ,Biology (General) ,QH301-705.5 - Abstract
Background: Hepatocellular carcinoma (HCC) represents a global health challenge. Effective biomarkers are required for an early diagnosis to improve the survival rates of HCC patients. Exonuclease 1 (EXO1) plays a significant role in the DNA repair and recombination mechanisms. This study aimed to investigate the diagnostic and prognostic roles of EXO1 in HCC.Methods: We analyzed the EXO1 expression levels in various cancers including HCC from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. RNA sequencing data were analyzed using the R packages to determine differentially expressed genes (DEGs) between high- and low-EXO1 expressing HCC tissues from the TCGA–LIHC database. A Spearman’s correlation analysis was performed to determine the association between EXO1 expression and immune cell infiltration, and immune checkpoint genes and TP53. MethSurv and CBioPortal databases were used to evaluate the DNA methylation changes and genetic alterations in the EXO1 gene. A logistic regression analysis was performed to determine the association between EXO1 expression and the clinicopathological characteristics of the HCC patients. The diagnostic and prognostic predictive values of EXO1 were evaluated using the Kaplan–Meier (K-M) survival curves, diagnostic receiver operating characteristic (ROC) curves, nomogram model, and Cox regression analysis.Results: EXO1 expression levels were significantly higher in the tumor tissues and serums of HCC patients compared to the corresponding controls. The DEGs associated with EXO1 were significantly enriched in the cell proliferation pathways. EXO1 expression levels significantly correlated with immune cell infiltration, immune checkpoint genes, and TP53 in the HCC tissues. The DNA methylation status in five CpG islands of the EXO1 gene was associated with the prognosis of HCC. EXO1 expression levels in the HCC tissues were associated with the tumor grades, alpha-fetoprotein (AFP) levels, and the tumor stages. Cox regression analysis showed that EXO1 was a potential independent risk factor for the overall survival (OS) and disease-specific survival (DSS) of HCC patients. ROC curve analysis showed that EXO1 expression levels accurately distinguished HCC tissues from the adjacent normal liver tissues.Conclusion: Our study demonstrated that EXO1 was a potential diagnostic and prognostic biomarker, and a promising therapeutic target in HCC.
- Published
- 2022
- Full Text
- View/download PDF
17. Genome Replication Is Associated With Release of Immunogenic DNA Waste
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Nadja Schubert, Tina Schumann, Elena Daum, Karolin Flade, Yan Ge, Lara Hagedorn, Winfried Edelmann, Luise Müller, Marc Schmitz, Gunnar Kuut, Veit Hornung, Rayk Behrendt, and Axel Roers
- Subjects
Trex1 ,type I interferon ,Exo1 ,replication ,cytosolic DNA ,interferonopathy ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Innate DNA sensors detect foreign and endogenous DNA to induce responses to infection and cellular stress or damage. Inappropriate activation by self-DNA triggers severe autoinflammatory conditions, including Aicardi-Goutières syndrome (AGS) that can be caused by defects of the cytosolic DNase 3’repair exonuclease 1 (TREX1). TREX1 loss-of-function alleles are also associated with systemic lupus erythematosus (SLE). Chronic activation of innate antiviral immunity in TREX1-deficient cells depends on the DNA sensor cGAS, implying that accumulating TREX1 DNA substrates cause the inflammatory pathology. Retrotransposon-derived cDNAs were shown to activate cGAS in TREX1-deficient neuronal cells. We addressed other endogenous sources of cGAS ligands in cells lacking TREX1. We find that induced loss of TREX1 in primary cells induces a rapid IFN response that requires ongoing proliferation. The inflammatory phenotype of Trex1-/- mice was partially rescued by additional knock out of exonuclease 1, a multifunctional enzyme providing 5’ flap endonuclease activity for Okazaki fragment processing and postreplicative ribonucleotide excision repair. Our data imply genome replication as a source of DNA waste with pathogenic potential that is efficiently degraded by TREX1.
- Published
- 2022
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18. DNA End Resection: Mechanism and Control.
- Author
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Cejka, Petr and Symington, Lorraine S.
- Abstract
DNA double-strand breaks (DSBs) are cytotoxic lesions that threaten genome integrity and cell viability. Typically, cells repair DSBs by either nonhomologous end joining (NHEJ) or homologous recombination (HR). The relative use of these two pathways depends on many factors, including cell cycle stage and the nature of the DNA ends. A critical determinant of repair pathway selection is the initiation of 5′→3′ nucleolytic degradation of DNA ends, a process referred to as DNA end resection. End resection is essential to create single-stranded DNA overhangs, which serve as the substrate for the Rad51 recombinase to initiate HR and are refractory to NHEJ repair. Here, we review recent insights into the mechanisms of end resection, how it is regulated, and the pathological consequences of its dysregulation. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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19. FANCD2-Associated Nuclease 1 Partially Compensates for the Lack of Exonuclease 1 in Mismatch Repair.
- Author
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Kratz, Katja, Artola-Borán, Mariela, Kobayashi-Era, Saho, Koh, Gene, Oliveira, Goncalo, Kobayashi, Shunsuke, Oliveira, Andreia, Xueqing Zou, Richter, Julia, Masataka Tsuda, Hiroyuki Sasanuma, Shunichi Takeda, Loizou, Joanna I., Sartori, Alessandro A., Nik-Zainal, Serena, and Jiricny, Josef
- Subjects
- *
EUKARYOTIC cells , *PHENOTYPES , *NUCLEASES , *DRUG resistance , *COLON cancer - Abstract
Germline mutations in the mismatch repair (MMR) genes MSH2, MSH6, MLH1, and PMS2 are linked to cancer of the colon and other organs, characterized by microsatellite instability and a large increase in mutation frequency. Unexpectedly, mutations in EXO1, encoding the only exonuclease genetically implicated in MMR, are not linked to familial cancer and cause a substantially weaker mutator phenotype. This difference could be explained if eukaryotic cells possessed additional exonucleases redundant with EXO1. Analysis of the MLH1 interactome identified FANCD2-associated nuclease 1 (FAN1), a novel enzyme with biochemical properties resembling EXO1. We now show that FAN1 efficiently substitutes for EXO1 in MMR assays and that this functional complementation is modulated by its interaction with MLH1. FAN1 also contributes to MMR in vivo; cells lacking both EXO1 and FAN1 have an MMR defect and display resistance to Nmethyl-N-nitrosourea (MNU) and 6-thioguanine (TG). Moreover, FAN1 loss amplifies the mutational profile of EXO1-deficient cells, suggesting that the two nucleases act redundantly in the same antimutagenic pathway. However, the increased drug resistance and mutator phenotype of FAN1/EXO1-deficient cells are less prominent than those seen in cells lacking MSH6 or MLH1. Eukaryotic cells thus apparently possess additional mechanisms that compensate for the loss of EXO1. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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20. EXO1 protects BRCA1-deficient cells against toxic DNA lesions.
- Author
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van de Kooij, Bert, Schreuder, Anne, Pavani, Raphael, Garzero, Veronica, Uruci, Sidrit, Wendel, Tiemen J., van Hoeck, Arne, San Martin Alonso, Marta, Everts, Marieke, Koerse, Dana, Callen, Elsa, Boom, Jasper, Mei, Hailiang, Cuppen, Edwin, Luijsterburg, Martijn S., van Vugt, Marcel A.T.M., Nussenzweig, André, van Attikum, Haico, and Noordermeer, Sylvie M.
- Subjects
- *
DNA damage , *DOUBLE-strand DNA breaks , *HOMOLOGOUS recombination , *EXONUCLEASES , *BRCA genes - Abstract
Inactivating mutations in the BRCA1 and BRCA2 genes impair DNA double-strand break (DSB) repair by homologous recombination (HR), leading to chromosomal instability and cancer. Importantly, BRCA1/2 deficiency also causes therapeutically targetable vulnerabilities. Here, we identify the dependency on the end resection factor EXO1 as a key vulnerability of BRCA1-deficient cells. EXO1 deficiency generates poly(ADP-ribose)-decorated DNA lesions during S phase that associate with unresolved DSBs and genomic instability in BRCA1-deficient but not in wild-type or BRCA2-deficient cells. Our data indicate that BRCA1/EXO1 double-deficient cells accumulate DSBs due to impaired repair by single-strand annealing (SSA) on top of their HR defect. In contrast, BRCA2-deficient cells retain SSA activity in the absence of EXO1 and hence tolerate EXO1 loss. Consistent with a dependency on EXO1-mediated SSA, we find that BRCA1 -mutated tumors show elevated EXO1 expression and increased SSA-associated genomic scars compared with BRCA1-proficient tumors. Overall, our findings uncover EXO1 as a promising therapeutic target for BRCA1-deficient tumors. [Display omitted] • EXO1 is essential for the survival of BRCA1- but not BRCA2-deficient cells • EXO1 loss induces S phase PAR signaling in both BRCA1- and BRCA2-deficient cells • EXO1 loss causes toxic DNA double-strand break accumulation in BRCA1-deficient cells • The double-strand break accumulation is caused by impaired single-strand annealing van de Kooij et al. identify loss of the exonuclease EXO1 as a vulnerability of BRCA1-deficient cells. Mechanistically, cells deficient for both BRCA1 and EXO1 suffer from unrepaired DNA double-stranded breaks due to the loss of two break repair pathways, homologous recombination, and single-strand annealing. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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21. DNA damage responses to loss of telomere integrity
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Carlos, A. R. and Tarsounas, M.
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572.8 ,Biology (medical sciences) ,DNA damage signalling ,Telomere integrity ,DNA damage response ,BRCA1 ,CtIP ,EXO1 - Abstract
Linear genomes end in characteristic structures consisting of repetitive DNA and proteins: the telomeres. These play two critical roles: on one hand they avoid the of loss of genetic information due to the incomplete replication of the chromosome ends and on the other, they provide capping structures for chromosome termini, differentiating them from double strand breaks. Telomeres contain specialized proteins (the shelterin complex), as well as proteins present elsewhere on the chromosomes (chromatin remodelling, DNA damage repair and response factors). Interestingly, several DNA damage factors are required for proper telomere maintenance, drawing a thin line between telomere protection and their recognition as broken DNA ends. Loss of telomere integrity has severe consequences for the cell, namely it can induce replicative senescence and cellular aging, or it can contribute to tumorigenesis. How telomeres are capped and how they are perceived by the cell when they become dysfunctional is essential for our understanding of the contribution of loss of telomere integrity to aging and disease. In order to unravel new factors involved in telomere maintenance, siRNA screens were performed. The optimization process has confirmed both telomeric foci and telomere dysfunction-induced foci (TIFs) as suitable readouts and the screens performed generated a list of potential candidate genes involved in telomere biology. Although some of the candidate genes tested in this work failed the validation process, other genes deserve further analysis. In addition this work also studied the role of several DNA damage factors at uncapped telomeres. Furthermore, BRCA1, CtIP and EXO1 were found to be critical for the formation of end-to-end fusions generated after TRF2 inactivation. The requirement of this proteins in this process, suggests that not only that not only the classical non-homologous end joining (C-NHEJ) pathway is active at TRF2-depelted telomeres, but emphasises the multiplicity of mechanisms that act to repair dysfunctional telomeres.
- Published
- 2013
22. Exonuclease 1 (EXO1) is a Potential Prognostic Biomarker and Correlates with Immune Infiltrates in Lung Adenocarcinoma.
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Zhou, Chang-shuai, Feng, Ming-tao, Chen, Xin, Gao, Yang, Chen, Lei, Li, Liang-dong, Li, De-heng, and Cao, Yi-qun
- Subjects
- *
PROGNOSIS , *CELL cycle regulation , *CANCER cell migration , *B cells , *BIOMARKERS - Abstract
Background: Exonuclease 1 (EXO1) has been identified to be highly expressed in different human malignancies, but its expression and prognostic role in lung adenocarcinoma (LUAD) remain unknown. Materials and Methods: Two independent cohorts extracted from public databases and one cohort from our center were analyzed in this study. Expression levels of EXO1 in LUAD tissues and paired para-cancer tissues were detected. The prognostic value of EXO1 in LUAD patients was evaluated in the three cohorts. Enrichment analyses were performed to explore the possible underlying biological pathways. Moreover, we also explored the correlations between EXO1 and tumor-infiltrating immune cells and evaluated the impact of EXO1 knock-down on the migration of lung cancer cells. Results: In this study, we found that EXO1 was highly expressed in LUAD tissues compared with para-cancerous tissues in public databases (p < 0.01), which was consistent with our data (p < 0.01). Survival analysis indicated that high expression of EXO1 was associated with poor prognosis in LUAD (p < 0.01). Enrichment analyses indicated that biological pathways like cell cycle regulation, DNA damage and repair, immune response, neuroactive ligand-receptor interaction, may be associated with EXO1 aberrant expression. Moreover, high expression of EXO1 was correlated with decreased infiltrating B cells (p < 0.01) and CD4+ T cells (p < 0.01) levels, and low infiltrating levels of B cells (p < 0.01) and dendritic cells (DCs) (p < 0.05) indicated poor overall survival (OS) in LUAD. Additionally, in vitro experiments suggested that knockdown of EXO1 may inhibit the migratory ability of lung cancer cells. Conclusion: In conclusion, EXO1 is a potential prognostic biomarker in LUAD, and correlates with infiltrating levels of immune cells in the tumor microenvironment. Further prospective validation of EXO1 in lung cancer is warranted. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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23. Evolution of Prime Editing Systems: Move Forward to the Treatment of Hereditary Diseases.
- Author
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Volodina OV, Fabrichnikova AR, Anuchina AA, Mishina OS, Lavrov AV, and Smirnikhina SA
- Abstract
The development of gene therapy using genome editing tools recently became relevant. With the invention of programmable nucleases, it became possible to treat hereditary diseases due to introducing targeted double strand break in the genome followed by homology directed repair (HDR) or non-homologous end-joining (NHEJ) reparation. CRISPR-Cas9 is more efficient and easier to use in comparison with other programmable nucleases. To improve the efficiency and safety of this gene editing tool, various modifications CRISPR-Cas9 basis were created in recent years, such as prime editing - in this system, Cas9 nickase is fused with reverse transcriptase and guide RNA, which contains a desired correction. Prime editing demonstrates equal or higher correction efficiency as HDR-mediated editing and much less off-target effect due to inducing nick. There are several studies in which prime editing is used to correct mutations in which researchers reported little or no evidence of off-target effects. The system can also be used to functionally characterize disease variants. However, prime editing still has several limitations that could be further improved. The effectiveness of the method is not yet high enough to apply it in clinical trials. Delivery of prime editors is also a big challenge due to their size. In the present article, we observe the development of the platform, and discuss the candidate proteins for efficiency enhancing, main delivery methods and current applications of prime editing., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)
- Published
- 2024
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24. Mutant huntingtin protein induces MLH1 degradation, DNA hyperexcision, and cGAS-STING-dependent apoptosis.
- Author
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Sun X, Liu L, Wu C, Li X, Guo J, Zhang J, Guan J, Wang N, Gu L, Yang XW, and Li GM
- Subjects
- Humans, Huntingtin Protein genetics, Huntingtin Protein metabolism, Mutant Proteins genetics, Nucleotidyltransferases genetics, DNA, Apoptosis genetics, MutL Protein Homolog 1 genetics, Huntington Disease genetics, Huntington Disease metabolism
- Abstract
Huntington's disease (HD) is an inherited neurodegenerative disorder caused by an expanded CAG repeat in the huntingtin ( HTT ) gene. The repeat-expanded HTT encodes a mutated HTT (mHTT), which is known to induce DNA double-strand breaks (DSBs), activation of the cGAS-STING pathway, and apoptosis in HD. However, the mechanism by which mHTT triggers these events is unknown. Here, we show that HTT interacts with both exonuclease 1 (Exo1) and MutLα (MLH1-PMS2), a negative regulator of Exo1. While the HTT-Exo1 interaction suppresses the Exo1-catalyzed DNA end resection during DSB repair, the HTT-MutLα interaction functions to stabilize MLH1. However, mHTT displays a significantly reduced interaction with Exo1 or MutLα, thereby losing the ability to regulate Exo1. Thus, cells expressing mHTT exhibit rapid MLH1 degradation and hyperactive DNA excision, which causes severe DNA damage and cytosolic DNA accumulation. This activates the cGAS-STING pathway to mediate apoptosis. Therefore, we have identified unique functions for both HTT and mHTT in modulating DNA repair and the cGAS-STING pathway-mediated apoptosis by interacting with MLH1. Our work elucidates the mechanism by which mHTT causes HD., Competing Interests: Competing interests statement:The authors declare no competing interest.
- Published
- 2024
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25. Variants in Homologous Recombination Genes EXO1 and RAD51 Related with Premature Ovarian Insufficiency.
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Wei Luo, Ting Guo, Guangyu Li, Ran Liu, Shidou Zhao, Meihui Song, Liangran Zhang, Shunxin Wang, Zi-Jiang Chen, Yingying Qin, Luo, Wei, Guo, Ting, Li, Guangyu, Liu, Ran, Zhao, Shidou, Song, Meihui, Zhang, Liangran, Wang, Shunxin, Chen, Zi-Jiang, and Qin, Yingying
- Subjects
MEIOSIS ,PREMATURE ovarian failure ,HAPLOIDY ,MENSTRUAL cycle ,HEREDITARY nonpolyposis colorectal cancer ,REPLICATION protein A ,RESEARCH ,GENETICS ,GENETIC mutation ,SEQUENCE analysis ,RESEARCH methodology ,EVALUATION research ,MEDICAL cooperation ,COMPARATIVE studies ,ENZYMES ,OVARIAN diseases ,ESTERASES - Abstract
Context: Premature ovarian insufficiency (POI) is characterized by cessation of menstruation before 40 years of age and elevated serum level of FSH (>25 IU/L). Recent studies have found a few causative genes responsible for POI enriched in meiotic recombination and DNA damage repair pathways.Objective: To investigate the role of variations in homologous recombination genes played in POI pathogenesis.Methods: The whole exome sequencing was performed in 50 POI patients with primary amenorrhea. Functional characterizations of the novel variants were carried out in budding yeast and human cell line.Results: We identified 8 missense variants in 7 homologous recombination genes, including EXO1, RAD51, RMI1, MSH5, MSH2, MSH6, and MLH1. The mutation p.Thr52Ser in EXO1 impaired the meiotic process of budding yeast and p.Glu68Gly in RAD51-altered protein localization in human cells, both of them impaired the efficiency of homologous recombination repair for DNA double-stranded breaks in human cells.Conclusions: Our study first linked the variants of EXO1 and RAD51 with POI and further highlighted the role of DNA repair genes in ovarian dysgenesis. [ABSTRACT FROM AUTHOR]- Published
- 2020
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26. 限制性外切酶Ⅰ(EXO1)的合成、克隆与表达.
- Author
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谢钰珍, 覃鸿妮, 张勇, and 赵文玮
- Subjects
- *
RECOMBINANT proteins , *AFFINITY chromatography , *SINGLE-stranded DNA , *DNA , *ESCHERICHIA , *GENE expression - Abstract
[Objective] The present paper aimed tl Exolease I(EX01) is a ultifunctionnal3’→5’exolease that was mainly used to digest single-stranded segments in double-stated DNA or RNA samples.Most of existing conimercialized EXO1 are induced from Escherichia coli.But low expression level and complex purification steps limit its application. The present paper aimed to synthesize sbcB gene, highly express in E.coli and purify the expressed product. [Method] The sbcB gene was synthesized by overlapping PCR and overexpressed by pET-30 a system in E.coli BL21(DE3), which was induced by IPTG, and the expression conditions were optimized to obtain the high expression of EXO1. The recombinant protein was purified by affinity chromatography and the enzyme activity was studide. [Result] The product EXO1 was consistent with the predicted 54.5 KD. Further functional verification also showed that EXO1 can digest single chains but can not digest double chains. The enzyme activity of purified interest protein was well. [Conclusion] EXO1 prepared by this method showed high yield, high purity, and was suitable for pre-treatment of PCR products in laboratory-sequencing. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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27. PARylation of BRCA1 limits DNA break resection through BRCA2 and EXO1
- Author
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Samuele, L., Roberto, Q., Sertic, S., and Pellicioli, A.
- Subjects
BRCA1/2 ,DSB resection ,DNA repair ,CP: Molecular biology ,Settore BIO/11 - Biologia Molecolare ,53BP1 barrier ,EXO1 ,PARP ,olaparib - Published
- 2023
28. Genetic dissection of crossover mutants defines discrete intermediates in mouse meiosis.
- Author
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Premkumar, Tolkappiyan, Paniker, Lakshmi, Kang, Rhea, Biot, Mathilde, Humphrey, Ericka, Destain, Honorine, Ferranti, Isabella, Okulate, Iyinyeoluwa, Nguyen, Holly, Kilaru, Vindhya, Frasca, Melissa, Chakraborty, Parijat, and Cole, Francesca
- Subjects
- *
MEIOSIS , *HOLLIDAY junctions , *CHROMOSOME segregation , *MICE , *DISSECTION - Abstract
Crossovers (COs), the exchange of homolog arms, are required for accurate chromosome segregation during meiosis. Studies in yeast have described the single-end invasion (SEI) intermediate: a stabilized 3′ end annealed with the homolog as the first detectible CO precursor. SEIs are thought to differentiate into double Holliday junctions (dHJs) that are resolved by MutLgamma (MLH1/MLH3) into COs. Currently, we lack knowledge of early steps of mammalian CO recombination or how intermediates are differentiated in any organism. Using comprehensive analysis of recombination in thirteen different genetic conditions with varying levels of compromised CO resolution, we infer CO precursors include asymmetric SEI-like intermediates and dHJs in mouse. In contrast to yeast, MLH3 is structurally required to differentiate CO precursors into dHJs. We verify conservation of aspects of meiotic recombination and show unique features in mouse, providing mechanistic insight into CO formation. [Display omitted] • Dissection of recombination in mouse spermatocytes defective for crossing over • EXO1 promotes MutLgamma endonuclease activity but is not fully required • Formation of single-end invasion-like and double Holliday junction intermediates • MLH3 plays a structural role to promote double Holliday junction formation Accurate chromosome segregation in meiosis requires crossovers, but we lack an understanding of how they form in mammals. Premkumar et al. dissect recombination in multiple crossover mutants to show unique and conserved mechanisms in mice. They find two distinct precursors and show that MLH3 is structurally required to form double Holliday junctions. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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29. Local unwinding of double-strand DNA ends by the MRX complex promotes Exo1 processing activity
- Author
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Elisa Gobbini, Jacopo Vertemara, and Maria Pia Longhese
- Subjects
double-strand break ,exo1 ,mrx ,resection ,s. cerevisiae ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Homologous recombination is initiated by nucleolytic degradation (resection) of DNA double-strand breaks (DSBs), which involves different nucleases including the Mre11-Rad50-Xrs2 (MRX) complex and the Exonuclease 1 (Exo1). The characterization of a novel mutation in Mre11 causing accelerated DSB resection has allowed to show that MRX facilitates DNA end processing by Exo1 through local unwinding of double-stranded DNA ends.
- Published
- 2018
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30. LncRNA CECR7 boosts hepatocellular carcinoma progression by recruiting RNA binding protein U2AF2 to enhance the stability of EXO1 mRNA.
- Author
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Zhao L, Zang Q, Liang G, and Yao X
- Abstract
Objective: As an important factor tumor regulator,long non-coding RNAs (lncRNAs) have aroused extensive attention via the diverse functional mechanisms that were associated with the pathological and physiological processes of HCC. Here, the main purpose of this study was to provide a clear understanding about the expression, functions and potential mechanism of lncRNA CECR7 (Cat Eye Syndrome Chromosome Region, Candidate 7) in HCC., Methods: RT-qPCR analysis and TCGA database analysis were applied to investigate the expression of CECR7 in HCC cell lines and tissues. Chi-squared Test was employed to explore the correlation between CECR7 expression and HCC clinicopathological features. Besides, Kaplan-Meier curves were constructed to test the effects of CECR7 expression on the prognosis of HCC patients. Transwell assays, MTT assay EdU assay and animal experiments were applied to explore the effects of CECR7 expression on HCC cells migration, invasion, and growth. Furthermore, RNA-seq analysis, luciferase reporter assay and mRNA decay rates assessment were utilized to investigate the mechanism whereby CECR7 regulated EXO1 mRNA. And, rescue experiments were used to determine whether EXO1 was an essential mediator for CECR7 to accelerate HCC cells migration, invasion, and growth., Results: CECR7 was determined to be significantly overexpressed in HCC cell lines and tissues. CECR7 expression was closely correlated with the tumor size, venous infiltration, TNM stage, 5-year overall survival and disease-free survival of HCC. And, CECR7 played a catalytic role in HCC cells migration, invasion, and growth. Furthermore, CECR7 enhanced the stability of EXO1 mRNA by recruiting RNA binding protein U2AF2. And, EXO1 was determined to be an essential mediator for CECR7 to accelerate HCC cells migration, invasion, and growth., Conclusion: In a word, our findings demonstrates that the cancer-promoting gene lncRNA CECR7 motivates HCC metastasis and growth through enhanced mRNA stability of EXO1 mediated by U2AF2, proposing a new insight for targeted therapy of HCC., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)
- Published
- 2023
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31. EXO1
- Author
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Choi, Sangdun, editor
- Published
- 2018
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32. EXO1 overexpression is associated with poor prognosis of hepatocellular carcinoma patients.
- Author
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Dai, Yaoyao, Tang, Zuxiong, Yang, Zongguo, Zhang, Lan, Deng, Qing, Zhang, Xiaofeng, Yu, Yongchun, Liu, Xing, and Zhu, Junfeng
- Abstract
The roles of exonuclease 1 (EXO1) in hepatocellular carcinoma (HCC) tumorigenesis and progression remain unclear. This study aimed to assess the prognostic value and therapeutic potential of EXO1 in HCC. Exo1 gene copy numbers were obtained from three Oncomine microarray datasets (n = 447). EXO1 mRNA expression was validated by semi-quantitative PCR and QuantiGene® 2.0 assays. Cell growth curve and colony formation were performed to asses the cell proliferation. Clonogenic assay, flow cytometry, and immunofluorescence were adopted to acess the effects of EXO1 knockdown and radiation on cell survival, cell cycle distribution and DNA repair. Western blots were performed to reveal the related mechanism. A significant copy number variation (CNV) of the Exo1 gene was found in HCC specimens in three separate sets of published microarray data. In the 143 cases treated by our team, EXO1 expression levels were elevated (86.71%, 124/143). In addition, EXO1 overexpression was correlated with larger tumor size (P = 0.002), increased lymph node metastasis (P=0.033) and lower Edmondson grade (P = 0.018). High EXO1 expression unfavorably affected overall survival (OS) (P = 0.009). Both univariate and multivariate Cox regression analyses identified EXO1 as an independent predictor of OS (univariate, P = 0.012; multivariate, P = 0.039). Silencing of EXO1 in vitro reduced cell proliferation. EXO1 knockdown further suppressed clonogenic cell survival, abrogated radiation-induced G2/M phase arrest, and enhanced γ-H2AX foci after exposure to irradiation. The accumulation of ataxiatelangiectasia mutated (ATM) might partially regulate the EXO1 related radiosensitivity. In summary, EXO1 could be a promising prognostic marker, with a potential therapeutic value in HCC. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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33. The MRX complex regulates Exo1 resection activity by altering DNA end structure.
- Author
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Gobbini, Elisa, Cassani, Corinne, Vertemara, Jacopo, Wang, Weibin, Mambretti, Fabiana, Casari, Erika, Sung, Patrick, Tisi, Renata, Zampella, Giuseppe, and Longhese, Maria Pia
- Subjects
- *
DOUBLE-strand DNA breaks , *RECOMBINANT DNA , *MOLECULAR dynamics , *NUCLEASES , *DNA structure - Abstract
Abstract: Homologous recombination is triggered by nucleolytic degradation (resection) of DNA double‐strand breaks (DSBs). DSB resection requires the Mre11‐Rad50‐Xrs2 (MRX) complex, which promotes the activity of Exo1 nuclease through a poorly understood mechanism. Here, we describe the Mre11‐R10T mutant variant that accelerates DSB resection compared to wild‐type Mre11 by potentiating Exo1‐mediated processing. This increased Exo1 resection activity leads to a decreased association of the Ku complex to DSBs and an enhanced DSB resection in G1, indicating that Exo1 has a direct function in preventing Ku association with DSBs. Molecular dynamics simulations show that rotation of the Mre11 capping domains is able to induce unwinding of double‐strand DNA (dsDNA). The R10T substitution causes altered orientation of the Mre11 capping domain that leads to persistent melting of the dsDNA end. We propose that MRX creates a specific DNA end structure that promotes Exo1 resection activity by facilitating the persistence of this nuclease on the DSB ends, uncovering a novel MRX function in DSB resection. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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34. Spatial separation between replisome‐ and template‐induced replication stress signaling.
- Author
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García‐Rodríguez, Néstor, Morawska, Magdalena, Wong, Ronald P., Daigaku, Yasukazu, and Ulrich, Helle D.
- Subjects
- *
REPLISOMES , *POLYMERASES , *DNA , *DNA replication , *DNA synthesis - Abstract
Abstract: Polymerase‐blocking DNA lesions are thought to elicit a checkpoint response via accumulation of single‐stranded DNA at stalled replication forks. However, as an alternative to persistent fork stalling, re‐priming downstream of lesions can give rise to daughter‐strand gaps behind replication forks. We show here that the processing of such structures by an exonuclease, Exo1, is required for timely checkpoint activation, which in turn prevents further gap erosion in S phase. This Rad9‐dependent mechanism of damage signaling is distinct from the Mrc1‐dependent, fork‐associated response to replication stress induced by conditions such as nucleotide depletion or replisome‐inherent problems, but reminiscent of replication‐independent checkpoint activation by single‐stranded DNA. Our results indicate that while replisome stalling triggers a checkpoint response directly at the stalled replication fork, the response to replication stress elicited by polymerase‐blocking lesions mainly emanates from Exo1‐processed, postreplicative daughter‐strand gaps, thus offering a mechanistic explanation for the dichotomy between replisome‐ versus template‐induced checkpoint signaling. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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35. Human Exonuclease 1 (EXO1) Regulatory Functions in DNA Replication with Putative Roles in Cancer
- Author
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Guido Keijzers, Daniela Bakula, Michael Angelo Petr, Nils Gedsig Kirkelund Madsen, Amanuel Teklu, Garik Mkrtchyan, Brenna Osborne, and Morten Scheibye-Knudsen
- Subjects
DNA repair ,double strand break repair ,exonuclease 1 ,EXO1 ,mismatch repair ,MMR ,NER ,nucleotide excision repair ,strand displacements ,TLS ,translesion DNA synthesis ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Human exonuclease 1 (EXO1), a 5′→3′ exonuclease, contributes to the regulation of the cell cycle checkpoints, replication fork maintenance, and post replicative DNA repair pathways. These processes are required for the resolution of stalled or blocked DNA replication that can lead to replication stress and potential collapse of the replication fork. Failure to restart the DNA replication process can result in double-strand breaks, cell-cycle arrest, cell death, or cellular transformation. In this review, we summarize the involvement of EXO1 in the replication, DNA repair pathways, cell cycle checkpoints, and the link between EXO1 and cancer.
- Published
- 2018
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36. Significant association of the EXO1 rs851797 polymorphism with clinical outcome of ovarian cancer.
- Author
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Rong Jiang, Pan Wang, Sheng Yin, Tingyan Shi, Rongyu Zang, Yuan Xu, and Xi Cheng
- Subjects
- *
EXONUCLEASES , *CANCER invasiveness , *OVARIAN cancer treatment , *GENETIC polymorphisms , *REGRESSION analysis - Abstract
Background: Exonuclease 1 (EXO1), one of DNA mismatch repair pathway genes, functions in maintaining genomic stability and affects tumor progression. We hypothesized that genetic variations in EXO1 may predict clinical outcomes in epithelial ovarian cancer (EOC). Methods: In this cohort study with 1,030 consecutive EOC patients, we genotyped four potentially functional polymorphisms in EXO1 by the Taqman assay and evaluated their associations with patients' survival. Results: Using multivariate Cox proportional hazards regression models, we found that rs851797AG/GG genotypes were significantly associated with recurrence and cancer death (HR =1.30 and 1.38, 95% CI =1.11-1.52 and 1.02-1.88, respectively). Kaplan-Meier survival estimates showed that patients who carried rs851797AG/GG genotypes had poorer progression-free survival and poorer overall survival, compared with rs851797AA genotype carriers (log-rank test, P=0.002 and 0.025, respectively). Moreover, patients with older age at menophania, advanced stage tumor, or being received incomplete cytoreduction were more likely to be recurrent and dead. Conclusion: EXO1 rs851797 polymorphism can predict the clinical outcomes in EOC patients. In addition, age at menophania, FIGO stage, and complete cytoreduction might be independently prognostic factors of ovarian cancer. Large studies with functional experiments are warranted to validate these findings. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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37. EXO1 suppresses double-strand break induced homologous recombination between diverged sequences in mammalian cells.
- Author
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Chen, Chun-Chin, Avdievich, Elena, Zhang, Yongwei, Zhang, Yu, Wei, Kaichun, Lee, Kyeryoung, Edelmann, Winfried, Jasin, Maria, and LaRocque, Jeannine R.
- Subjects
- *
DOUBLE-strand DNA breaks , *DNA damage , *DNA mismatch repair , *PROTEINS , *EMBRYONIC stem cells - Abstract
DNA double-strand breaks (DSBs) can be repaired through several mechanisms, including homologous recombination (HR). While HR between identical sequences is robust in mammalian cells, HR between diverged sequences is suppressed by DNA mismatch-repair (MMR) components such as MSH2. Exonuclease I (EXO1) interacts with the MMR machinery and has been proposed to act downstream of the mismatch recognition proteins in mismatch correction. EXO1 has also been shown to participate in extensive DSB end resection, an initial step in the HR pathway. To assess the contribution of EXO1 to HR in mammalian cells, DSB-inducible reporters were introduced into Exo1 −/− mouse embryonic stem cells, including a novel GFP reporter containing several silent polymorphisms to monitor HR between diverged sequences. Compared to HR between identical sequences which was not clearly affected, HR between diverged sequences was substantially increased in Exo1 −/− cells although to a lesser extent than seen in Msh2 −/− cells. Thus, like canonical MMR proteins, EXO1 can restrain aberrant HR events between diverged sequence elements in the genome. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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38. PHF11 promotes DSB resection, ATR signaling, and HR.
- Author
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Yi Gong, Naofumi Handa, Kowalczykowski, Stephen C., and de Lange, Titia
- Subjects
- *
NUCLEOTIDE sequencing , *NUCLEOTIDE analysis , *NUCLEOTIDE sequence , *SEQUENCE alignment , *CELL death - Abstract
Resection of double-strand breaks (DSBs) plays a critical role in their detection and appropriate repair. The 3' ssDNA protrusion formed through resection activates the ATR-dependent DNA damage response (DDR) and is required for DSB repair by homologous recombination (HR). Here we report that PHF11 (plant homeodomain finger 11) encodes a previously unknown DDR factor involved in 5' end resection, ATR signaling, and HR. PHF11 was identified based on its association with deprotected telomeres and localized to sites of DNA damage in S phase. Depletion of PHF11 diminished the ATR signaling response to telomere dysfunction and genome-wide DNA damage, reduced end resection at sites of DNA damage, resulted in compromised HR and misrejoining of S-phase DSBs, and increased the sensitivity to DNA-damaging agents. PHF11 interacted with the ssDNA-binding protein RPA and was found in a complex with several nucleases, including the 5' dsDNA exonuclease EXO1. Biochemical experiments demonstrated that PHF11 stimulates EXO1 by overcoming its inhibition by RPA, suggesting that PHF11 acts (in part) by promoting 5' end resection at RPA-bound sites of DNA damage. These findings reveal a role for PHF11 in DSB resection, DNA damage signaling, and DSB repair. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
39. PARylation of BRCA1 limits DNA break resection through BRCA2 and EXO1.
- Author
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Lodovichi, Samuele, Quadri, Roberto, Sertic, Sarah, and Pellicioli, Achille
- Abstract
The nucleolytic processing (resection) of a DNA double-strand break (DSB) is a critical step to repair the lesion by homologous recombination (HR). PARylation, which is the attachment of poly(ADP-ribose) (PAR) units to specific targets by PAR polymerases (PARPs), regulates many steps of HR, including resection. Here, we show that preventing PARylation of the oncosuppressor BRCA1 induces hyper-resection of DSBs through BRCA2 and the EXO1 nuclease. Upon expression of the unPARylatable variant of BRCA1, we observe a reduced 53BP1-RIF1 barrier for resection accompanied by an increase in the recruitment of the RAD51 recombinase. Similar results are observed when cells are treated with the clinically approved PARP inhibitor olaparib. We propose that PARylation of BRCA1 is important to limit the formation of excessively extended DNA filaments, thereby reducing illegitimate chromosome rearrangements. Our results shed light on molecular aspects of HR and on the mechanisms of PARP inhibitor treatment. [Display omitted] • PARP1 inhibition leads to over-resection of Cas9-induced DNA double-strand breaks • Downregulation of BRCA1 or BRCA2 abrogates excessive PARPi-dependent DNA resection • Expression of unPARylable BRCA1 variant induces over-resection of Cas9-induced DSBs • BRCA2 and EXO1 promotes DSB over-resection in unPARylable BRCA1 cells Lodovichi et al. show that BRCA1 PARylation limits excessive resection of DNA double-strand breaks (DSBs). Inhibition of PARP1 or expression of an unPARylable variant of BRCA1 leads to over-resection of DSBs via the BRCA2/EXO1 axis, promoting RAD51-dependent DNA repair. These findings elucidate the mechanism of PARP inhibition in cancer therapy. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
40. Long-range DNA end resection supports homologous recombination by checkpoint activation rather than extensive homology generation.
- Author
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Kimble MT, Johnson MJ, Nester MR, and Symington LS
- Subjects
- DNA Breaks, Double-Stranded, DNA Repair, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, Homologous Recombination, Exodeoxyribonucleases genetics, Exodeoxyribonucleases metabolism, DNA metabolism, RecQ Helicases metabolism, Saccharomyces cerevisiae Proteins genetics, Saccharomyces cerevisiae Proteins metabolism
- Abstract
Homologous recombination (HR), the high-fidelity mechanism for double-strand break (DSB) repair, relies on DNA end resection by nucleolytic degradation of the 5'-terminated ends. However, the role of long-range resection mediated by Exo1 and/or Sgs1-Dna2 in HR is not fully understood. Here, we show that Exo1 and Sgs1 are dispensable for recombination between closely linked repeats, but are required for interchromosomal repeat recombination in Saccharomyces cerevisiae . This context-specific requirement for long-range end resection is connected to its role in activating the DNA damage checkpoint. Consistent with this role, checkpoint mutants also show a defect specifically in interchromosomal recombination. Furthermore, artificial activation of the checkpoint partially restores interchromosomal recombination to exo1∆ sgs1∆ cells. However, cell cycle delay is insufficient to rescue the interchromosomal recombination defect of exo1∆ sgs1∆ cells, suggesting an additional role for the checkpoint. Given that the checkpoint is necessary for DNA damage-induced chromosome mobility, we propose that the importance of the checkpoint, and therefore long-range resection, in interchromosomal recombination is due to a need to increase chromosome mobility to facilitate pairing of distant sites. The need for long-range resection is circumvented when the DSB and its repair template are in close proximity., Competing Interests: MK, MJ, MN, LS No competing interests declared, (© 2023, Kimble et al.)
- Published
- 2023
- Full Text
- View/download PDF
41. Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ.
- Author
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Tomomi Aida, Shota Nakade, Tetsushi Sakuma, Yayoi Izu, Ayu Oishi, Keiji Mochida, Harumi Ishikubo, Takako Usami, Hidenori Aizawa, Takashi Yamamoto, and Kohichi Tanaka
- Subjects
- *
GENE cassettes , *FUNCTIONAL analysis , *ZYGOTES , *HOMOLOGY (Biochemistry) , *EXONUCLEASES - Abstract
Background: Although CRISPR/Cas enables one-step gene cassette knock-in, assembling targeting vectors containing long homology arms is a laborious process for high-throughput knock-in. We recently developed the CRISPR/Cas-based precise integration into the target chromosome (PITCh) system for a gene cassette knock-in without long homology arms mediated by microhomology-mediated end-joining. Results: Here, we identified exonuclease 1 (Exo1) as an enhancer for PITCh in human cells. By combining the Exo1 and PITCh-directed donor vectors, we achieved convenient one-step knock-in of gene cassettes and floxed allele both in human cells and mouse zygotes. Conclusions: Our results provide a technical platform for high-throughput knock-in. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
42. Exonuclease 1 and its versatile roles in DNA repair.
- Author
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Keijzers, Guido, Liu, Dekang, and Rasmussen, Lene Juel
- Subjects
- *
EXONUCLEASES , *IMMUNOGLOBULINS , *GENETIC mutation , *DNA repair , *DNA structure , *POST-translational modification - Abstract
Exonuclease 1 (EXO1) is a multifunctional 5′ → 3′ exonuclease and a DNA structure-specific DNA endonuclease. EXO1 plays roles in DNA replication, DNA mismatch repair (MMR) and DNA double-stranded break repair (DSBR) in lower and higher eukaryotes and contributes to meiosis, immunoglobulin maturation, and micro-mediated end-joining in higher eukaryotes. In human cells, EXO1 is also thought to play a role in telomere maintenance. Mutations in the humanEXO1gene correlate with increased susceptibility to some cancers. This review summarizes recent studies on the enzymatic functions and biological roles of EXO1, its possible protective role against cancer and aging, and regulation of EXO1 by posttranslational modification. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
43. EXO1 Plays a Carcinogenic Role in Hepatocellular Carcinoma and is related to the regulation of FOXP3
- Author
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Er-lei Zhang, Bin-yong Liang, Keshuai Dong, Zun-yi Zhang, Xiaoping Chen, Zhiyong Huang, and Guang Yang
- Subjects
Gene knockdown ,Nuclease ,biology ,Oncogene ,FOXP3 ,Hepatocellular carcinoma ,Chemistry ,Cell growth ,EXO1 ,Metastasis ,Exonuclease 1 ,Transcriptional regulation ,Oncology ,Cancer research ,biology.protein ,Transcription factor ,Research Paper - Abstract
Exonuclease 1 (EXO1), a member of the RAD2 nuclease family, was first described as possessing 5' to 3' nuclease activity and 5' structure-specific endonuclease activity. Here, we show that EXO1 is significantly upregulated in HCC tumor tissues and that high EXO1 expression is significantly correlated with liver cirrhosis. We further demonstrate that EXO1 knockdown decreases proliferation and colony forming abilities of HCC cells in vitro and tumorigenicity in vivo, as well as decreases migration and invasive capabilities of HCC cells. Alternatively, EXO1 overexpression significantly increases the proliferation, colony forming ability, and migration and invasive capabilities of HCC cells in vitro. Additionally, we truncated a region upstream of the transcription start site (TSS) of EXO1 and used the region with the strongest transcriptional activity to predict that the transcription factor FOXP3 can bind to the EXO1 promoter. Bioinformatics analysis found that FOXP3 was positively correlated with EXO1 and luciferase reporter assays and RT-PCR confirmed that FOXP3 could enhance the transcriptional activity of EXO1. CCK-8 assays showed that depletion of FOXP3 further reduces cell proliferation ability after knocking down of EXO1 in vitro. Taken together, our findings indicate that EXO1 acts as an oncogene in HCC and its expression level is related to FOXP3 activity.
- Published
- 2020
44. Breast cancer-associated SNP rs72755295 is a cis-regulatory variation for human EXO1
- Author
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Qiang Shi, Xing-Yuan Yao, Hong-Yan Wang, Ya-Jie Li, Xin-Xin Zhang, and Chang Sun
- Subjects
Breast cancer ,expression regulation ,Genetics ,rs72755295 ,rs4149909 ,EXO1 ,Molecular Biology - Abstract
Breast cancer is the most common malignant tumor in women. A previous genome-wide association study reports that rs72755295, a SNP locating at intron of EXO1 (exonuclease 1), is associated with breast cancer. Due to the complete linkage disequilibrium between rs72755295 and rs4149909, a nonsynonymous mutation for EXO1, rs4149909 is supposed to be the causal SNP. Since EXO1 is overexpressed in breast carcinoma samples, we hypothesized that the genetic variations in this locus might confer breast cancer risk by regulating EXO1 expression. To substantiate this, a functional genomics study was performed. The dual luciferase assay indicated that G of rs72755295 presents significantly higher relative enhancer activity than A, thus verifying that this SNP can influence gene expression in breast cell. Through chromosome conformation capture it was disclosed that the enhancer containing rs72755295 can interact with the EXO1 promoter. RNA-seq analysis indicated that EXO1 expression is dependent on the rs72755295 genotype. By chromatin immunoprecipitation, the transcription factor PAX6 (paired box 6) was recognized to bind the region spanning rs72755295. In electrophoretic mobility shift assay, G of rs72755295 displays obviously higher binding affinity with nuclear protein than A. Our results indicated that rs72755295 is a cis-regulatory variation for EXO1 and might confer breast cancer risk besides rs4149909.
- Published
- 2021
45. FANCD2-associated nuclease 1 partially compensates for the lack of Exonuclease 1 in mismatch repair
- Author
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Gene Koh, Josef Jiricny, Serena Nik-Zainal, Hiroyuki Sasanuma, Alessandro A. Sartori, Saho Kobayashi-Era, Goncalo Oliveira, Katja Kratz, Andreia Oliveira, Julia Richter, Shunichi Takeda, Shunsuke Kobayashi, Masataka Tsuda, Mariela Artola-Borán, Joanna I. Loizou, Xueqing Zou, and University of Zurich
- Subjects
Exonuclease ,Methylnitronitrosoguanidine ,congenital, hereditary, and neonatal diseases and abnormalities ,exonuclease ,DNA repair ,610 Medicine & health ,EXO1 ,DNA Mismatch Repair ,Cell Line ,Avian Proteins ,03 medical and health sciences ,Exonuclease 1 ,0302 clinical medicine ,Germline mutation ,FAN1 ,PMS2 ,Animals ,Humans ,Amino Acid Sequence ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Endodeoxyribonucleases ,Thionucleosides ,biology ,Base Sequence ,Guanosine ,10061 Institute of Molecular Cancer Research ,MLH1 ,Cell Biology ,mutational signature ,MSH6 ,Multifunctional Enzymes ,digestive system diseases ,Cell biology ,mismatch repair ,Exodeoxyribonucleases ,HEK293 Cells ,MSH2 ,030220 oncology & carcinogenesis ,Mutation ,biology.protein ,570 Life sciences ,DNA mismatch repair ,Chickens ,Research Article - Abstract
Germline mutations in the mismatch repair (MMR) genes MSH2, MSH6, MLH1, and PMS2 are linked to cancer of the colon and other organs, characterized by microsatellite instability and a large increase in mutation frequency. Unexpectedly, mutations in EXO1, encoding the only exonuclease genetically implicated in MMR, are not linked to familial cancer and cause a substantially weaker mutator phenotype. This difference could be explained if eukaryotic cells possessed additional exonucleases redundant with EXO1. Analysis of the MLH1 interactome identified FANCD2-associated nuclease 1 (FAN1), a novel enzyme with biochemical properties resembling EXO1. We now show that FAN1 efficiently substitutes for EXO1 in MMR assays and that this functional complementation is modulated by its interaction with MLH1. FAN1 also contributes to MMR in vivo; cells lacking both EXO1 and FAN1 have an MMR defect and display resistance to N-methyl-N-nitrosourea (MNU) and 6-thioguanine (TG). Moreover, FAN1 loss amplifies the mutational profile of EXO1-deficient cells, suggesting that the two nucleases act redundantly in the same antimutagenic pathway. However, the increased drug resistance and mutator phenotype of FAN1/EXO1-deficient cells are less prominent than those seen in cells lacking MSH6 or MLH1. Eukaryotic cells thus apparently possess additional mechanisms that compensate for the loss of EXO1., Cellular and Molecular Biology, 41 (9), ISSN:0145-5680, ISSN:1165-158X
- Published
- 2021
46. Rif1 and Exo1 regulate the genomic instability following telomere losses.
- Author
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Xue, Yuan, Marvin, Marcus E., Ivanova, Iglika G., Lydall, David, Louis, Edward J., and Maringele, Laura
- Subjects
- *
TELOMERES , *GENOMICS , *CELLULAR aging , *DNA damage , *CELL proliferation - Abstract
Telomere attrition is linked to cancer, diabetes, cardiovascular disease and aging. This is because telomere losses trigger further genomic modifications, culminating with loss of cell function and malignant transformation. However, factors regulating the transition from cells with short telomeres, to cells with profoundly altered genomes, are little understood. Here, we use budding yeast engineered to lack telomerase and other forms of telomere maintenance, to screen for such factors. We show that initially, different DNA damage checkpoint proteins act together with Exo1 and Mre11 nucleases, to inhibit proliferation of cells undergoing telomere attrition. However, this situation changes when survivors lacking telomeres emerge. Intriguingly, checkpoint pathways become tolerant to loss of telomeres in survivors, yet still alert to new DNA damage. We show that Rif1 is responsible for the checkpoint tolerance and proliferation of these survivors, and that is also important for proliferation of cells with a broken chromosome. In contrast, Exo1 drives extensive genomic modifications in survivors. Thus, the conserved proteins Rif1 and Exo1 are critical for survival and evolution of cells with lost telomeres. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
47. Mechanism and regulation of DNA end resection in eukaryotes.
- Author
-
Symington, Lorraine S.
- Subjects
- *
DNA repair , *EUKARYOTES , *DOUBLE-strand polymers , *GENETIC recombination , *DNA damage , *NUCLEASES - Abstract
The repair of DNA double-strand breaks (DSBs) by homologous recombination (HR) is initiated by nucleolytic degradation of the 5′-terminated strands in a process termed end resection. End resection generates 3′-single-stranded DNA tails, substrates for Rad51 to catalyze homologous pairing and DNA strand exchange, and for activation of the DNA damage checkpoint. The commonly accepted view is that end resection occurs by a two-step mechanism. In the first step, Sae2/CtIP activates the Mre11–Rad50–Xrs2/Nbs1 (MRX/N) complex to endonucleolytically cleave the 5′-terminated DNA strands close to break ends, and in the second step Exo1 and/or Dna2 nucleases extend the resected tracts to produce long 3′-ssDNA-tailed intermediates. Initiation of resection commits a cell to repair a DSB by HR because long ssDNA overhangs are poor substrates for non-homologous end joining (NHEJ). Thus, the initiation of end resection has emerged as a critical control point for repair pathway choice. Here, I review recent studies on the mechanism of end resection and how this process is regulated to ensure the most appropriate repair outcome. [ABSTRACT FROM PUBLISHER]
- Published
- 2016
- Full Text
- View/download PDF
48. Influence of a single-nucleotide polymorphism of the DNA mismatch repair-related gene exonuclease-1 (rs9350) with prostate cancer risk among Chinese people.
- Author
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Zhang, Yiming, Li, Pengju, Xu, Abai, Chen, Jie, Ma, Chao, Sakai, Akiko, Xie, Liping, Wang, Lei, Na, Yanqun, Kaku, Haruki, Xu, Peng, Jin, Zhong, Li, Xiezhao, Guo, Kai, Shen, Haiyan, Zheng, Shaobo, Kumon, Hiromi, Liu, Chunxiao, and Huang, Peng
- Abstract
In this study, we aimed to identify the influence of exonuclease 1 ( EXO1) single-nucleotide polymorphism rs9350, which is involved in DNA mismatch repair, on prostate cancer risk in Chinese people. In our hospital-based case-control study, 214 prostate cancer patients and 253 cancer-free control subjects were enrolled from three hospitals in China. Genotyping for rs9350 was performed by the SNaPshot method using peripheral blood samples. Consequently, a significantly higher prostate cancer risk was observed in patients with the CC genotype [odds ratio (OR) = 1.678, 95 % confidence interval (CI) = 1.130-2.494, P = 0.010] than in those with the CT genotype. Further, the CT/TT genotypes were significantly associated with increased prostate cancer risk (adjusted OR = 1.714, 95 % CI = 1.176-2.500, P = 0.005), and the C allele had a statistically significant compared with T allele ( P = 0.009) of EXO1 (rs9350). Through stratified analysis, significant associations were revealed for the CT/TT genotype in the subgroup with diagnosis age >72 (adjusted OR = 1.776, 95 % CI = 1.051-3.002, P = 0.032) and in patients with localized disease subgroup (adjusted OR = 1.798, 95 % CI = 1.070-3.022, P = 0.027). In addition, we observed that patients with prostate-specific antigen (PSA) levels of ≤10 ng/mL were more likely to have the CT/TT genotypes than those with PSA levels of >10 ng/mL ( P = 0.006). For the first time, we present evidence that the inherited EXO1 polymorphism rs9350 may have a substantial influence on prostate cancer risk in Chinese people. We believe that the rs9350 could be a useful biomarker for assessing predisposition for and early diagnosis of prostate cancer. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
49. Variation analysis of EXO1 gene in Chinese patients with premature ovarian failure.
- Author
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Su, Shizhen, Han, Ting, Ma, Bowen, Li, Weiping, Qin, Yingying, Zhao, Shidou, and Chen, Zi-Jiang
- Subjects
- *
PREMATURE ovarian failure , *EXONUCLEASES , *DNA repair , *MEIOSIS , *PUBLIC health , *GENETIC mutation - Abstract
Exonuclease 1 (EXO1) is required for both DNA repair and meiosis. Inactivation of EXO1 gene in mice leads to infertility. This study aimed to investigate whether variants in the EXO1 gene contribute to human premature ovarian failure (POF). The coding region of EXO1 was sequenced in 186 Han Chinese patients with non-syndromic POF. No plausible mutation was detected. The results suggest that mutations in the coding region of EXO1 may not be responsible for POF in Han Chinese women. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
50. Poly(ADP-ribose)-binding promotes Exo1 damage recruitment and suppresses its nuclease activities.
- Author
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Cheruiyot, Abigael, Paudyal, Sharad C., Kim, In-Kwon, Sparks, Melanie, Ellenberger, Tom, Piwnica-Worms, Helen, and You, Zhongsheng
- Subjects
- *
DEOXYRIBOSE , *DNA , *NUCLEIC acids , *DEOXYURIDINE triphosphate , *ESCHERICHIA nucleic acids , *GENE transfection - Abstract
Exonuclease 1 (Exo1) has important roles in DNA metabolic transactions that are essential for genome maintenance, telomere regulation and cancer suppression. However, the mechanisms for regulating Exo1 activity in these processes remain incompletely understood. Here, we report that Exo1 activity is regulated by a direct interaction with poly(ADP-ribose) (PAR), a prominent posttranslational modification at the sites of DNA damage. This PAR-binding activity promotes the early recruitment of Exo1 to sites of DNA damage, where it is retained through an interaction with PCNA, which interacts with the C-terminus of Exo1. The effects of both PAR and PCNA on Exo1 damage association are antagonized by the 14-3-3 adaptor proteins, which interact with the central domain of Exo1. Although PAR binding inhibits both the exonuclease activity and the 5′ flap endonuclease activity of purified Exo1, the pharmacological blockade of PAR synthesis does not overtly affect DNA double-strand break end resection in a cell free Xenopus egg extract. Thus, the counteracting effects of PAR on Exo1 recruitment and enzymatic activity may enable appropriate resection of DNA ends while preventing unscheduled or improper processing of DNA breaks in cells. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
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