JOURNAL OF CELL BIOLOGY
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2025
·Vol. 225
·期 1
·e202501091
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IF 6.4
Gao Kun,Shi Ruiya,Xu Chenying,Mao Zhaoyang,Guo Changying,Jin Liang
DNA damage repair is vital for maintaining genetic stability and integrity of cells that encounter DNA-damaging agents. So far, a series of multifunctional proteins and long noncoding RNAs (lncRNAs) have been demonstrated to participate in the DNA damage response (DDR). However, our current understanding of detailed mechanisms of DNA damage repair remains limited. Herein, we report that lncRNA EGFR-AS1 is functionally involved in DDR in both non–small-cell lung cancer cells and noncancerous cells. Using DNA repair reporter, we found that EGFR-AS1 overexpression significantly enhances the efficiency of both the classical nonhomologous end-joining and homologous recombination pathways. Through the lncRNA interactome, we identified a set of DNA repair factors, including the canonical DNA damage sensor PARP1 and NAD+ supplier NMNAT1. Upon DNA damage, DNA-activated PARP1 binds to EGFR-AS1 and forms a ternary complex with NMNAT1, promoting NAD+ utilization and poly(ADP-ribosyl)ation (PARylation) of PARP1. Additionally, EGFR-AS1 also facilitates displacing PARP1 from the sites of damaged DNA. Our findings demonstrate a lncRNA-associated PARP1 activation and displacement in DDR and highlight the potential of EGFR-AS1 as a target for cancer therapy.
研究领域:
分子生物学
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肿瘤学
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遗传学
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细胞生物学
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转化医学
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DOI
PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Suling Yang,Yaliang Huang,Tingting Yang,Jingze Li,Jiaxin Tian,Lin Liu
Poly(ADP-ribose)polymerase-1 (PARP1) could be activated by binding to nucleic acids with specific sequences, thus catalyzing the poly-ADP-ribosylation (PARylation) of target proteins including PARP1 itself. Most of the previously reported electrochemical methods for the determination of PARP1 were relied on the electrostatic interactions, which required the pre-immobilization of DNA on an electrode for the capture of PARP1. Herein, we reported an “immobilization−free” electrochemical strategy for the assays of PARP1 on the basic of avidin-biotin interaction. Once PARP1 was activated by binding with the specific double-stranded DNA (dsDNA) in a homogeneous solution, the biotinylated nicotinamide adenine dinucleotide (biotin-NAD + ) was transferred onto PARP1, resulting in the formation of biotinylated PAR polymers. The resulting biotinylated PAR polymers were then captured by a neutravidin (NA)-modified electrode through avidin-biotin interactions. The rich biotin moieties in the PAR polymers allowed for the capture of NA-modified silver nanoparticles (NA-AgNPs) through the avidin-biotin interactions. The surface-tethered AgNPs produced a well-defined electrochemical signal due to the characteristic solid-state Ag/AgCl process. The “immobilization−free”, electrostatic interaction-independent electrochemical biosensor exhibited low background current, high sensitivity, and good stability. It has achieved the determination of PARP1 with a detection limit down to 0.7 mU. The biosensor was further applied to determine the inhibition efficiency of potential inhibitors with a satisfactory result. This method shows promising potential applications in PARP1-related clinical diagnosis and drug discovery.
研究领域:
生物化学
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医学诊断
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纳米技术
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产品关联
DOI
PubMed
高影响因子
MOLECULAR CELL
·
2025
·
IF 16.6
Lu-ning Qin,Ting Wu,Xue-ting Zhen,Yun-long Zhao,Jie-shi Zhou,Shan-bin Cheng,Kai-wen Wang,Shuai Qiao,Ding-zhi Huang,Heng Zhang,Tao Sun
The link between extrachromosomal DNA (ecDNA) and tumors has been well established, and its role in cancer is of increasing interest. While ecDNA is thought to originate from genomic instability, the molecular mechanisms driving DNA end ligation during ecDNA formation and the regulatory factors controlling its selective gene packaging remain unresolved. Here, using the multi-layer perceptron model, a series of imaging strategies in human cancer cells, and clinical chip verification, we demonstrate that ecDNA biogenesis depends on transcription factor Yin Yang 1 (YY1)-mediated DNA looping coupled with religation catalyzed by DNA ligase 3 (Lig3), a mechanism that extends existing models. Notably, PARylation-dependent acidic microenvironments mediated by the Lig3-YY1 complex play a critical role in the formation of Z-DNA, which potentially facilitates the fusion-religation process to drive ecDNA biogenesis. Furthermore, our findings establish PARP inhibitors as specific agents for ecDNA-targeted strategies in cancer therapy.
研究领域:
分子生物学
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遗传学
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癌症生物学
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生物信息学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2018
·Vol. 143
·期 11
·2501-2507
·
IF 3.864
Yong Liu,Xiaolin Xu,Haitang Yang,Ensheng Xu,Shuangshuang Wu,Wei Wei,Jin Chen
Poly(ADP-ribose) polymerase-1 (PARP-1) is a highly conserved nuclear enzyme, which binds tightly to damaged DNA and plays a key role in DNA repair, recombination, proliferation, and genomic stability. However, due to the poor electrochemical and optical activity of PARP-1 and its product PAR, only a few studies on its activity detection method have been reported. Herein, we report a simple and sensitive colorimetric strategy to monitor PARP-1 activity based on enzyme-initiated auto-PARylation-controlled aggregation of hemin-graphene nanocomposites (H-GNs). PARP, activated by dsDNA, catalyzed its substrate nicotinamide adenine dinucleotide (NAD+) to polymerize as a poly(ADP-ribose) polymer (PAR). PAR possesses several negative charges, and its charge density is twice that of a single-stranded DNA, which greatly impacts the dispersibility of H-GNs; due to their peroxidase-like catalytic activities, H-GNs can catalyze the chromogenic reaction of TMB and H2O2. As a result, in the presence of different PARP-1 activities, the supernatant of the corresponding solution contained different amounts of dispersed H-GNs and showed different colors after the chromogenic reaction that could be discerned easily by the absorbance or the color changes of the solution. The method was simple, sensitive, and reliable. The proposed method displays a linear range from 0.05 to 1 U with a detection limit of 0.03 U. In addition, this new method has been successfully applied to detect PARP-1 activity in human serum and different cancer cells and evaluate PARP-1 inhibitors.
研究领域:
生物化学
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分子生物学
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癌症研究
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纳米技术
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DOI
PubMed
高影响因子
EUROPEAN JOURNAL OF PHARMACOLOGY
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2024
·
IF 4.2
Ya-Ling Chen,Yi Wang,Qiu-Yu Fang,Tong Wang,Cong Chen,Tong-Yao Gao,Ming Wu,Wei-Ping Zhang,Yun-Bi Lu
Poly (ADP-ribose) polymerase-1 (PARP-1) activity significantly increases during cerebral ischemia/reperfusion. PARP-1 is an NAD + -consumption enzyme. PARP-1 hyperactivity causes intracellular NAD + deficiency and bioenergetic collapse, contributing to neuronal death. Besides, the powerful trigger of PARP-1 causes the catalyzation of poly (ADP-ribosyl)ation (PARylation), a posttranslational modification of proteins. Here, we found that PARP-1 was activated in the ischemic brain tissue during middle-cerebral-artery occlusion and reperfusion (MCAO/R) for 24 h, and PAR accumulated in the neurons in mice. Using immunoprecipitation, Western blotting, liquid chromatography-mass spectrometry, and 3D-modeling analysis, we revealed that the activation of PARP-1 caused PARylation of hexokinase-1 and lactate dehydrogenase-B, which, therefore, caused the inhibition of these enzyme activities and the resulting cell energy metabolism collapse. PARP-1 inhibition significantly reversed the activity of hexokinase and lactate dehydrogenase, decreased infarct volume, and improved neuronal deficiency. PARP-1 inhibitor combined with pyruvate further alleviated MCAO/R-induced ischemic brain injury in mice. As such, we conclude that PARP-1 inhibitor alleviates neuronal death partly by inhibiting the PARylation of metabolic-related enzymes and reversing metabolism reprogramming during cerebral ischemia/reperfusion injury in mice. PARP-1 inhibitor combined with pyruvate might be a promising therapeutic approach against brain ischemia/reperfusion injury.
研究领域:
神经科学
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药理学
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分子生物学
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
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2016
·
IF 7.145
Yuanyuan Xu,Li Liu,Zhaoyin Wang,Zhihui Dai
A stable and reusable electrochemical biosensor for the label-free detection of poly(ADP-ribose) polymerase (PARP) is designed in this work. C-kit-1, a thiol-modified G-quadruplex oligonucleotide, is first self-assembled on a gold electrode surface. The G-quadruplex structure of c-kit-1 can specifically tether and activate PARP, resulting in the generation of negatively charged poly(ADP-ribose) polymer (PAR). On the basis of electrostatic attraction, PAR facilitates the surface accumulation of positively charged electrochemical signal molecules. Through the characterization of electrochemical signal molecules, the label-free quantification of PARP is simply implemented. On the basis of the proposed method, selective quantification of PARP can be achieved over the linear range from 0.01 to 1 U with a calculated detection limit of 0.003U. Further studies also demonstrate the applicability of the proposed method to biosamples revealing the broad potential in practical applications. Furthermore, inhibitor of PARP has also been detected with this biosensor. Meanwhile, benefited from self-assembly on solid surface, this biosensor possesses two important features, i.e., reusability and stability, which are desirable in related biosensors.
研究领域:
生物化学
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分子生物学
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分析化学
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产品关联
DOI
PubMed
高影响因子
Biosensors-Basel
·
2023
·Vol. 13
·期 7
·723
·
IF 5.4
Fengli Gao,Gang Liu,Yishu Qiao,Xiuwen Dong,Lin Liu
This work reports the development of a fluorescence method for the detection of poly(ADP-ribose) polymerase-1 (PARP1), in which a phenylboronic acid-modified fluorescein isothiocyanate dye (FITC-PBA) was used to recognize the formed poly(ADP-ribose) (PAR) polymer. The detection system was designed by conjugating recombinant streptavidin (rSA) with PARP1-specific double-stranded DNA (dsDNA) through streptavidin–biotin interaction. Capture of PARP1 via rSA–biotin–dsDNA allowed for the poly-ADP-ribosylation (PARylation) of both rSA and PARP1 in a homogeneous solution. The resulting rSA–biotin–dsDNA/PAR conjugates were then captured and separated via the commercialized nitrilotriacetic acid–nickel ion-modified magnetic bead (MB-NTA-Ni) through the interaction between NTA–Ni on MB surface and oligohistidine (His6) tag in rSA. The PAR polymer could capture the dye of FITC-PBA through the borate ester interaction between the boronic acid moiety in PBA and thecis-diol group in ribose, thus causing a decrease in fluorescence signal. The PARylation of streptavidin and the influence of steric hindrance on PARylation efficiency were confirmed using reasonable detection strategies. The method showed a wide linear range (0.01~20 U) and a low detection limit (0.01 U). This work should be valuable for the development of novel biosensors for the detection of poly(ADP-ribose) polymerases and diol-containing species.Keywords:streptavidin;nitrilotriacetic acid;magnetic bead;poly(ADP-ribose) polymerase;boronic acid;immobilization-free
研究领域:
生物化学
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分析化学
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分子生物学
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生物技术
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DOI
PubMed
高影响因子
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