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PARP2 activity is regulated by PARP1 in response to DNA alkylating agent mitomycin C. Arabidopsis plants, including (A) parp1-1 or (B) parp1-2 knockout alleles of PARP1, were grown on MS plates supplemented with 0, 20 and 40 μM of mitomycin C (MMC) for two weeks. Total proteins were extracted, separated by SDS-PAGE and analyzed by immunoblotting with anti-PAR antibody. Equivalent loading of lanes was verified using Ponceau S stain. Upper and middle panels of (B) are same blot, showing immunoblot signal after longer and shorter exposure times respectively. Similar results obtained in two separate experiments.
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Poly (ADP-ribose) polymerases (PARPs) catalyze the transfer of multiple poly(ADP-ribose) units onto target proteins. Poly(ADP-ribosyl)ation plays a crucial role in a variety of cellular processes including, most prominently, auto-activation of PARP at sites of DNA breaks to activate DNA repair processes. In humans, PARP1 (the founding and most char...
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... activity is regulated by PARP1 in response to DNA alkylating agent mitomycin C In further work to characterize the role of PARPs in plant DNA damage responses, wild-type and parp mutant plants were treated with mitomycin C to induce DNA cross-linking [51] and the level of poly(ADP-ribosyl)ated proteins was then monitored. As shown in Fig 6, mitomycin C caused increased PARP activity in wild-type plants. As with the bleomycin and γ-ray experi- ments (Figs 3 and 4), almost no poly(ADP-ribosyl)ation activity was detected after mitomycin C treatment in parp2-1 mutant plants (Fig 6). ...
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... shown in Fig 6, mitomycin C caused increased PARP activity in wild-type plants. As with the bleomycin and γ-ray experi- ments (Figs 3 and 4), almost no poly(ADP-ribosyl)ation activity was detected after mitomycin C treatment in parp2-1 mutant plants (Fig 6). However, increased rather than decreased abun- dance of poly(ADP-ribosyl)ation was observed in parp1 mutants, in separate experiments with either the parp1-1 or parp1-2 alleles, in response to mitomycin C (Fig 6). ...
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... with the bleomycin and γ-ray experi- ments (Figs 3 and 4), almost no poly(ADP-ribosyl)ation activity was detected after mitomycin C treatment in parp2-1 mutant plants (Fig 6). However, increased rather than decreased abun- dance of poly(ADP-ribosyl)ation was observed in parp1 mutants, in separate experiments with either the parp1-1 or parp1-2 alleles, in response to mitomycin C (Fig 6). This is unlike the poly (ADP-ribosyl)ation behavior of the same mutants in response to bleomycin or γ-irradiation (Fig 2). ...
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... is unlike the poly (ADP-ribosyl)ation behavior of the same mutants in response to bleomycin or γ-irradiation (Fig 2). The mitomycin C-induced increase in poly(ADP-ribosyl)ation caused by mutation of PARP1 was eliminated if PARP2 was also mutated (parp1-1parp2-1 and parp1-2parp2-1 dou- ble mutants, Fig 6). This interesting finding suggests that loss of PARP1 can in some situations lead to elevated PARP2 activity. ...
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Purpose:
Poly(ADP-ribose) polymerase 1 (PARP1) is necessary for single-strand break (SSB) repair by sensing DNA breaks and facilitating DNA repair through poly ADP-ribosylation of several DNA-binding and repair proteins. Inhibition of PARP1 results in collapsed DNA replication fork and double-strand breaks (DSBs). Accumulation of DSBs goes beyond...
PARP1 and PARP2 are implicated in the synthesis of poly(ADP-ribose) (PAR) after detection of DNA damage. The specificity of
PARP1 and PARP2 interaction with long DNA fragments containing single- and/or double-strand breaks (SSBs and DSBs) have been
studied using atomic force microscopy (AFM) imaging in combination with biochemical approaches. Our d...
Breaks in DNA strands recruit the protein PARP1 and its paralogue PARP2 to modify histones and other substrates through the addition of mono- and poly(ADP-ribose) (PAR)1–5. In the DNA damage responses, this post-translational modification occurs predominantly on serine residues6–8 and requires HPF1, an accessory factor that switches the amino acid...
Poly(ADP‐ribosyl)ation is predominantly catalyzed by Poly(ADP‐ribose) polymerase 1 (PARP1) in response to DNA damage, mediating the DNA repair process to maintain genomic integrity. Single‐strand (SSB) and double‐strand (DSB) DNA breaks are bona fide stimulators of PARP1 activity. However, PAR‐mediated PARP1 regulation remains unexplored. Here, we...
Citations
... The initiation of the TMEJ repair pathway is believed to start with recognition of the DSB via PARP1 (Chen et al., 1994 P ;Doucet-Chabeaud et al., 2001 P ;Jia et al., 2013 P ). PARP1 is a broadly conserved protein across eukaryotes (Citarelli et al., 2010 PO ;Song et al., 2015) PO that searches for the DNA break via a rapid 'monkey bar' mechanism (Rudolph et al., 2018 O (Mosler et al., 2022 O ). TMEJ was reported to be active in the S and G 2 phases of the cell cycle (Sfeir and Symington, 2015 O ;Dutta et al., 2017 O ), and a recent study on human cell lines showed that TMEJ is specifically activated in the M phase when c-NHEJ and HR pathways are attenuated (Brambati et al., 2023 O ). ...
A tool for precise, target-specific, efficient, and affordable genome editing is a dream for many researchers, from those who conduct basic research to those who use it for applied research. Since 2012, we have tool that almost fulfils such requirements; it is based on clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) systems. However, even CRISPR/Cas has limitations and obstacles that might surprise its users. In this review, we focus on the most frequently used variant, CRISPR/Cas9 from Streptococcus pyogenes, and highlight key factors affecting its mutagenesis outcomes: (i) factors affecting the CRISPR/Cas9 activity, such as the effect of the target sequence, chromatin state, or Cas9 variant, and how long it remains in place after cleavage; and (ii) factors affecting the follow-up DNA repair mechanisms including mostly the cell type and cell cycle phase, but also, for example, the type of DNA ends produced by Cas9 cleavage (blunt/staggered). Moreover, we note some differences between using CRISPR/Cas9 in plants, yeasts, and animals, as knowledge from individual kingdoms is not fully transferable. Awareness of these factors can increase the likelihood of achieving the expected results of plant genome editing, for which we provide detailed guidelines.
... PARylation has been confirmed to play essential roles in numerous cellular processes, including DNA repair, transcription regulation, chromatin modification, and ribosome biogenesis (Gibson and Kraus, 2012;Bock et al., 2015;Dasovich and Leung, 2023). Moreover, protein PARylation has implications in plant immunity against different pathogens (Feng et al., 2015;Song et al., 2015). For example, disrupting AtPARPs or AtPARGs modifies Arabidopsis responses to biotic and abiotic stresses (Adams-Phillips et al., 2010;Feng et al., 2015;Song et al., 2015, Yao et al., 2021. ...
... Moreover, protein PARylation has implications in plant immunity against different pathogens (Feng et al., 2015;Song et al., 2015). For example, disrupting AtPARPs or AtPARGs modifies Arabidopsis responses to biotic and abiotic stresses (Adams-Phillips et al., 2010;Feng et al., 2015;Song et al., 2015, Yao et al., 2021. However, in filamentous fungi, PrpA, a putative PARP homolog in Aspergillus nidulans, functions early in the DNA damage response (Semighini et al., 2006). ...
Poly(ADP-ribosyl)ation (PARylation), catalyzed by poly(ADP-ribose) polymerases (PARPs) and hydrolyzed by poly(ADP-ribose) glycohydrolase (PARG), is a kind of post-translational protein modification that is involved in various cellular processes in fungi, plants, and mammals. However, the function of PARPs in plant pathogenic fungi remains unknown. The present study investigated the roles and mechanisms of FonPARP1 in watermelon Fusarium wilt fungus Fusarium oxysporum f. sp. niveum (Fon). Fon has a single PARP FonPARP1 and one PARG FonPARG1. FonPARP1 is an active PARP and contributes to Fon pathogenicity through regulating its invasive growth within watermelon plants, while FonPARG1 is not required for Fon pathogenicity. A serine/threonine protein kinase, FonKin4, was identified as a FonPARP1-interacting partner by LC–MS/MS. FonKin4 is required for vegetative growth, conidiation, macroconidia morphology, abiotic stress response and pathogenicity of Fon. The S_TKc domain is sufficient for both enzyme activity and pathogenicity function of FonKin4 in Fon. FonKin4 phosphorylates FonPARP1 in vitro to enhance its poly(ADP-ribose) polymerase activity; however, FonPARP1 does not PARylate FonKin4. These results establish the FonKin4-FonPARP1 phosphorylation cascade that positively contributes to Fon pathogenicity. The present study highlights the importance of PARP-catalyzed protein PARylation in regulating the pathogenicity of Fon and other plant pathogenic fungi.
... PARP2 is a major active enzyme in Arabidopsis, ensuring the viability of plants challenged by IR. In parallel, it restricts the growth of phytopathogenic bacteria (Song et al., 2015). Transcription factor SOG1 regulates the majority of the genes induced by γ-rays and promotes DNA repair. ...
In nature, plants are simultaneously exposed to different abiotic (e.g., heat, drought, and salinity) and biotic (e.g., bacteria, fungi, and insects) stresses. Climate change and anthropogenic pressure are expected to intensify the frequency of stress factors. Although plants are well equipped with unique and common defense systems protecting against stressors, they may compromise their growth and development for survival in such challenging environments. Ionizing radiation is a peculiar stress factor capable of causing clustered damage. Radionuclides are both naturally present on the planet and produced by human activities. Natural and artificial radioactivity affects plants on molecular, biochemical, cellular, physiological, populational, and transgenerational levels. Moreover, the fitness of pests, pathogens, and symbionts is concomitantly challenged in radiologically contaminated areas. Plant responses to artificial acute ionizing radiation exposure and laboratory-simulated or field chronic exposure are often discordant. Acute or chronic ionizing radiation exposure may occasionally prime the defense system of plants to better tolerate the biotic stress or could often exhaust their metabolic reserves, making plants more susceptible to pests and pathogens. Currently, these alternatives are only marginally explored. Our review summarizes the available literature on the responses of host plants, biotic factors, and their interaction to ionizing radiation exposure. Such systematic analysis contributes to improved risk assessment in radiologically contaminated areas.
... However, PARP2 and PARP3 have a more compact structure compared to PARP1, featuring only a short N-terminal extension from the WGR domain ( Figure 1A,B) [16]. Functionally, PARP2 is considered the primary PAR-forming PARP following PARP1 and can compensate for the loss of PARP1 in vivo [29,30]. Hence, the PARPi approved by the US Food and Drug Administration (FDA) ( Figure 1C, Table 1) for clinical application primarily targets PARP1 and PARP2, taking advantage of their crucial roles in DNA repair processes. ...
... However, PARP2 and PARP3 have a more compact structure compared to PARP1, featuring only a short N-terminal extension from the WGR domain (Figure 1A,B) [16]. Functionally, PARP2 is considered the primary PAR-forming PARP following PARP1 and can compensate for the loss of PARP1 in vivo [29,30]. Hence, the PARPi approved by the US Food and Drug Administration (FDA) ( Figure 1C, Table 1) for clinical application primarily targets PARP1 and PARP2, taking advantage of their crucial roles in DNA repair processes. ...
Simple Summary
Gastric cancer (GC) is a highly aggressive malignancy affecting the digestive system and characterized by notable variations in its nature. Although survival rates have observed some enhancements, the prognosis remains grim, and mortality rates remain high. There is an immediate demand for innovative treatment strategies. Recent investigations have identified frequent mutations in genes responsible for repairing DNA damage in GC patients, highlighting the potential of PARP inhibitors (PARPi) as a promising therapeutic option. This article offers a comprehensive examination of the progress and hurdles in the development of PARPi for treating GC.
Abstract
Gastric cancer (GC) is a common and aggressive cancer of the digestive system, exhibiting high aggressiveness and significant heterogeneity. Despite advancements in improving survival rates over the past few decades, GC continues to carry a worrisome prognosis and notable mortality. As a result, there is an urgent need for novel therapeutic approaches to address GC. Recent targeted sequencing studies have revealed frequent mutations in DNA damage repair (DDR) pathway genes in many GC patients. These mutations lead to an increased reliance on poly (adenosine diphosphate-ribose) polymerase (PARP) for DNA repair, making PARP inhibitors (PARPi) a promising treatment option for GC. This article presents a comprehensive overview of the rationale and development of PARPi, highlighting its progress and challenges in both preclinical and clinical research for treating GC.
... PARPs are DNA damage sensors that bind to DNA SSBs or DSBs and attract damage repair machinery. PARP2, rather than PARP1, has a role in basal defenses against bacterial pathogens (Song et al., 2015). The molecular mechanisms underlying the interactions between DDR signaling and immune responses to the viral pathogen are still elusive. ...
... The mac3a_mac3b double mutants showed susceptibility to bacterial pathogens like Pst DC3000, PsmES4326, avrRps4, and avrPphB, suggesting that MAC3A and MAC3B are related to basal and R protein-mediated defense pathways (Monaghan et al., 2009). However, the atm-2_atr-2 double mutants were more susceptible to bacterial pathogens compared to the single mutants, whereas sog101 plants were more susceptible to fungal pathogen compared to the atm-2 and atr-2 single mutant plants (Ogita et al., 2018;Song et al., 2015). These results implied that DDR defense responses to bacterial pathogens are mainly ATM/ATR-dependent (Ogita et al., 2018); whereas, DDR defense responses to fungal pathogens are SOG1-dependent. ...
... The DDR responses to pathogens were activated by various factors depending on the pathogen's lifestyle (Camborde et al., 2019). PARPs are key DNA damage sensors for chromatin modifications and cell death (Gibson and Kraus, 2012;Song et al., 2015). The PARPs accumulate in response to genotoxic stress for DNA repair at the damage sites (Gibson and Kraus, 2012;Song et al., 2015). ...
Plants are challenged by various pathogens throughout their lives, such as bacteria, viruses, fungi, and insects; consequently, they have evolved several defense mechanisms. In addition, plants have developed localized and systematic immune responses due to biotic and abiotic stress exposure. Animals are known to activate DNA damage responses (DDRs) and DNA damage sensor immune signals in response to stress, and the process is well studied in animal systems. However, the links between stress perception and immune response through DDRs remain largely unknown in plants. To determine whether DDRs induce plant resistance to pathogens, Arabidopsis plants were treated with bleomycin, a DNA damage-inducing agent, and the replication levels of viral pathogens and growth of bacterial pathogens were determined. We observed that DDR-mediated resistance was specifically activated against viral pathogens, including turnip crinkle virus (TCV). DDR increased the expression level of pathogenesis-related ( PR ) genes and the total salicylic acid (SA) content and promoted mitogen-activated protein kinase signaling cascades, including the WRKY signaling pathway in Arabidopsis. Transcriptome analysis further revealed that defense- and SA-related genes were upregulated by DDR. The atm-2atr-2 double mutants were susceptible to TCV, indicating that the main DDR signaling pathway sensors play an important role in plant immune responses. In conclusion, DDRs activated basal immune responses to viral pathogens.
... 49,50,52 PARP1 is the most abundant member of the PARP family in eukaryotes, which accounts for almost 90% of cellular poly(ADP-ribose) polymerase activity in response to DNA damage. 49,53 Although the basal enzymatic activity of PARP1 is negligible, the enzymatic activity is stimulated dramatically under conditions of DNA damage and cellular stress. [54][55][56] PARP1 plays a crucial role in maintaining genomic stability, transcription, cellular energy metabolism, and cell death. ...
... With A DET predicted to code for a prontein that enables DNA repair, namely, poly [ADP-RIBOSE] polymerase 3-like, displayed clearly enhanced expression upon priming. A similar protein found in Arabidopsis thaliana is related to protection against stress caused by gamma radiation [32]. Therefore, it may be argued that this protein plays a role during priming in the protection of DNA and, thus, contributes to successful germination under drought stress. ...
Solanum paniculatum L. belongs to the Solanaceae family and has the ability to grow and develop under unfavorable environmental conditions such as drought and salt stress, acid soils and soils poor in nutrients. The present work aimed to analyze S. paniculatum seed transcriptome associated with induced tolerance to drought stress by osmopriming. Seeds subjected to osmopriming (−1.0 MPa) displayed a higher germination and normal seedling percentage under drought stress when compared with unprimed seeds. RNA-seq transcriptome profiles of osmoprimed and unprimed seeds were determined and the potential proteins involved in the drought tolerance of S. paniculatum were identified. From the 34,640 assembled transcripts for both osmoprimed and unprimed seeds, only 235 were differentially expressed and, among these, 23 (10%) transcripts were predicted to code for proteins potentially involved in response to stress, response to abiotic stimulus and response to chemical. The possible mechanisms by which these stress-associated genes may confer tolerance to osmoprimed Solanum paniculatum seeds to germinate under water deficit was discussed and may help to find markers for the selection of new materials belonging to the Solanaceae family that are more tolerant to stress during and following germination.
... With regards to plant PARP1 and PARP2, although assumptions concerning their functions have often been inferred by analogy with animal PARPs, the plant-specific roles of these proteins have become apparent [1,2]. In particular, plant PARP1 and PARP2 proteins have been linked to plant responses to biotic and abiotic stresses [1,17,19]. ...
The nucleolus and Cajal bodies (CBs) are sub-nuclear domains with well-known roles in RNA metabolism and RNA-protein assembly. However, they also participate in other important aspects of cell functioning. This study uncovers a previously unrecognised mechanism by which these bodies and their components regulate host defences against pathogen attack. We show that the CB protein coilin interacts with poly(ADP-ribose) polymerase 1 (PARP1), redistributes it to the nucleolus and modifies its function, and that these events are accompanied by substantial increases in endogenous concentrations of salicylic acid (SA), activation of SA-responsive gene expression and callose deposition leading to the restriction of tobacco rattle virus (TRV) systemic infection. Consistent with this, we also find that treatment with SA subverts the negative effect of the pharmacological PARP inhibitor 3-aminobenzamide (3AB) on plant recovery from TRV infection. Our results suggest that PARP1 could act as a key molecular actuator in the regulatory network which integrates coilin activities as a stress sensor for virus infection and SA-mediated antivirus defence.
... PARylation plays an important role in a broad array of cellular processes, including DNA damage detection and repair, cell division and death, chromatin modification, and transcriptional and translational regulation [1][2][3][4][5][6][7][8][9]. An increasing number of studies in plants show that PARylation is involved in several biological processes, including circadian rhythms and responses to abiotic and biotic stresses [10][11][12][13][14][15][16]. ...
Poly(ADP-ribosyl)lation (PARylation) is a posttranslational modification that plays an important role in a variety of biological processes in both animals and plants. Identification of PARylated substrates is the key to elucidating the regulatory mechanism of PARylation. Several approaches have been developed to identify PARylated substrates over the past decade; however, a reliable and efficient method is needed to demonstrate PARylated proteins. Here, we report a simple and sensitive assay of PARylated proteins using a clickable 6-alkyne-NAD+ analog. The 6-alkyne-NAD+ is incorporated into substrate proteins in the in vitro PARylation assay. The labeled proteins are covalently captured by disulfide azide agarose beads through copper-catalyzed azide-alkyne cycloaddition (CuAAC), cleaved under reducing conditions, and analyzed by immunoblotting. The covalent bonds between the PARylated proteins and azide beads allow high stringent washing to eliminate nonspecific binding. Furthermore, the disulfide linker permits efficient cleavage and recovery of highly enriched PARylated proteins. Therefore, this approach can detect proteins that undergo PARylation at very low levels.Key wordsPoly(ADP-ribosyl)ationPARPsClick chemistry6-alkyne-NAD+ analog
... ERF115 activity is needed when stem cells are damaged [44]. RAD51 and BRCA1 encode two DNA repair regulators that are involved in the repair of double-stranded DNA breaks, and the expression of PARP2 is induced by ionizing radiation and radiomimetic drugs [45][46][47]. As shown in Figure 4F, the expression levels of all four genes were elevated after exposure to FB1. number of dead cells increased with a longer incubation ( Figure 4A and Supplemental Figure S1). ...
Fumonisin B1 is a mycotoxin that is structurally analogous to sphinganine and sphingosine and inhibits the biosynthesis of complex sphingolipids by repressing ceramide synthase. Based on the connection between FB1 and sphingolipid metabolism, FB1 has been widely used as a tool to explore the multiple functions of sphingolipids in mammalian and plant cells. The aim of this work was to determine the effect of sphingolipids on primary root development by exposing Arabidopsis (Arabidopsis thaliana) seedlings to FB1. We show that FB1 decreases the expression levels of several PIN-FORMED (PIN) genes and the key stem cell niche (SCN)-defining transcription factor genes WUSCHEL-LIKE HOMEOBOX5 (WOX5) and PLETHORAs (PLTs), resulting in the loss of quiescent center (QC) identity and SCN maintenance, as well as stunted root growth. In addition, FB1 induces cell death at the root apical meristem in a non-cell-type-specific manner. We propose that sphingolipids play a key role in primary root growth through the maintenance of the root SCN and the amelioration of cell death in Arabidopsis.
