• Title/Summary/Keyword: DNA damage tolerance

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Chronological Switch from Translesion Synthesis to Homology-Dependent Gap Repair In Vivo

  • Fujii, Shingo;Isogawa, Asako;Fuchs, Robert P.
    • Toxicological Research
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    • v.34 no.4
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    • pp.297-302
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    • 2018
  • Cells are constantly exposed to endogenous and exogenous chemical and physical agents that damage their genome by forming DNA lesions. These lesions interfere with the normal functions of DNA such as transcription and replication, and need to be either repaired or tolerated. DNA lesions are accurately removed via various repair pathways. In contrast, tolerance mechanisms do not remove lesions but only allow replication to proceed despite the presence of unrepaired lesions. Cells possess two major tolerance strategies, namely translesion synthesis (TLS), which is an error-prone strategy and an accurate strategy based on homologous recombination (homology-dependent gap repair [HDGR]). Thus, the mutation frequency reflects the relative extent to which the two tolerance pathways operate in vivo. In the present paper, we review the present understanding of the mechanisms of TLS and HDGR and propose a novel and comprehensive view of the way both strategies interact and are regulated in vivo.

Interferon-Stimulated Gene 15 in the Control of Cellular Responses to Genotoxic Stress

  • Jeon, Young Joo;Park, Jong Ho;Chung, Chin Ha
    • Molecules and Cells
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    • v.40 no.2
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    • pp.83-89
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    • 2017
  • Error-free replication and repair of DNA are pivotal to organisms for faithful transmission of their genetic information. Cells orchestrate complex signaling networks that sense and resolve DNA damage. Post-translational protein modifications by ubiquitin and ubiquitin-like proteins, including SUMO and NEDD8, are critically involved in DNA damage response (DDR) and DNA damage tolerance (DDT). The expression of interferon-stimulated gene 15 (ISG15), the first identified ubiquitin-like protein, has recently been shown to be induced under various DNA damage conditions, such as exposure to UV, camptothecin, and doxorubicin. Here we overview the recent findings on the role of ISG15 and its conjugation to target proteins (e.g., p53,$ {\Delta}Np63{\alpha}$, and PCNA) in the control of cellular responses to genotoxic stress, such as the inhibition of cell growth and tumorigenesis.

Antioxidative and Probiotic Properties of Lactobacillus gasseri NLRI-312 Isolated from Korean Infant Feces

  • Kim, H.S.;Jeong, S.G.;Ham, J.S.;Chae, H.S.;Lee, J.M.;Ahn, C.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.9
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    • pp.1335-1341
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    • 2006
  • We selected a Lactobacillus spp. from Korean healthy infant feces based upon their antioxidant activity. This strain was identified as Lactobacillus gasseri by 16S rDNA sequencing, and named Lactobacillus gasseri NLRI-312. In the present study, we investigate the protective effect of this strain on the $H_2O_2$ induced damage to cellular membrane lipid and DNA in Jurkat cells. To estimate the extent of cellular lipid peroxidation inhibition, MDA (malondialdehyde) was measured, and DNA damage was tested by the comet assay. We also examined probiotic properties including tolerance to acid and bile, antibiotic resistance. From the results obtained, the supplementation of Jurkat cells with NLRI-312 decreased in DNA damage, while no effect was shown on MDA decrease. In probiotic properties, this strain was resistance to both acid and bile, showed considerably higher survival when incubated in pH 2 or 1% bile salts (w/v). We concluded that the NLRI-312 could be used as potential probiotic bacteria, with the effect of reducing DNA damage induced by $H_2O_2$.

Nucleus-DNA Damage and Different Response of Plant Cells to Paraquat in Relation to Enzyme Activity of Superoxide Dismutase. (Superoxide dismutase의 활성차이에 따른 식물세포의 paraquat에 대한 반응과 핵 DNA 손상 검정)

  • 권순태;이명현;오세명;정도철;김길웅
    • Journal of Life Science
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    • v.14 no.4
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    • pp.614-619
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    • 2004
  • This study was undertaken to investigate the different responses of cultured plant cells to paraquat treatment and nucleus-DNA damage in relation to enzyme activity of superoxide dismutase (SOD). Furthermore, this study was also carried out to understand the antioxidative mechanism of plant cells to environmental stress. We selected two different species of plant cultured cells, Ipomoea batatas as high-SOD species and Lonicera japonica as low-SOD species. The total activity and specific activity of SOD in a chlorophyllous cell of I. batatas were 3,736 unit/gㆍfresh weight and 547 unit/mgㆍprotein, respectively, and those in L. japonica were 23 unit/gㆍfresh weight and 13 unit/mgㆍprotein, respectively SOD activity in chlorophyllous I. batatas cells reached its maximum level at 10 to 15 days after subculture, whereas that in L. japonica remained at a very low SOD level during the whole period of subculture. In comparison to L. japonica, I. batatas, a high-SOD species, showed high tolerance to paraquat 10 and 50 mg/l treatment in terms of cell viability and electrolyte leakage. Based on the result of comet assay, the nucleus-DNA damage of two species by paraquat 50 mg/l treatment was not significantly different. However, I. batatas cells repaired their damaged DNA more effectively than the cells of the low-SOD species, L. japonica.

Effect of long term treatment of aqueous extract of Enicostemma littorale in Type 2 diabetic patients

  • Mansuri, Mustakim M;Goyal, Bhoomika R;Upadhyay, Umesh M;Sheth, Jayesh;Goyal, Ramesh K
    • Advances in Traditional Medicine
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    • v.9 no.1
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    • pp.39-48
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    • 2009
  • We have evaluated the effect of long term treatment of Enicostemma littorale (E. littorale) in type 2 diabetic patients taking pills of aqueous extract of E. littorale regularly as a complimentary medicine for at least 9 months. The effects of E. littorale on glycemic control, lipid profile, cardiac function and DNA damage in these patients were compared with those who had not been regular in taking E. littorale but regular in taking other conventional anti-diabetics. Our data suggest that, E. littorale can maintain normal blood glucose, serum insulin, serum triglycerides levels of type 2 diabetic patients if taken regularly. E. littorale also improves insulin sensitivity, and normalize disturbed lipogram and elevated creatinine levels, thereby produces beneficial effect in preventing cardiovascular complications and may preserve the kidney function. The finding that E. littorale also prevents DNA damage suggest a long term effect in diabetic patients. E. littorale thus can be considered as safe supplementary therapy for a long term and effective management of type 2 diabetic patients.

Overexpression of NtROS2a gene encoding cytosine DNA demethylation enhances drought tolerance in transgenic rice (시토신 탈메틸화 관련 NtROS2a 유전자 도입 형질전환벼의 건조스트레스 내성 증진)

  • Choi, Jang Sun;Lee, In Hye;Cho, Yong-Gu;Jung, Yu Jin;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.376-382
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    • 2016
  • DNA methylation regulations gene expression, thus having pivotal roles in a myriad of physiological and pathological processes. In this study, the morphology and stress tolerance of transgenic rice overexpressing NtROS2a were determined. Transgenic plants exhibited less and shorter lateral shoots. Under various treatments, rice overexpressing NtROS2a showed alleviation of damage symptoms with higher survival rate. After drought and re-watering treatment, transgenic rice seedlings restored their normal growth. However, wild type plants could not be rescued. These findings indicate that overexpression of NtROS2a gene in rice seedlings can increase their tolerance to drought stresses.

Prevention of UV-induced Skin Damage by Activation of Tumor Suppressor Genes p53 and $p14^{ARF}$

  • Petersen, R.;John, S.;Lueder, M.;Borchert, S.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.338-351
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    • 2003
  • UV radiation is the most dangerous stress factor among permanent environmental impacts on human skin. Consequences of UV exposure are aberrant tissue architecture, alterations in skin cells including functional changes. Nowadays new kinds of outdoor leisure-time activities and changing environmental conditions make the question of sun protection more important than ever. It is necessary to recognize that self-confident consumers do not consider to change their way of life, they demand modern solutions on the basis of new scientific developments. In the past one fundamental principle of cosmetics was the use of physical and organic filter systems against damaging UV-rays. Today new research results demonstrate that natural protecting cell mechanisms can be activated. Suitable biological actives strongly support the protection function not from the surface but from the inside of the cell. A soy seed preparation (SSP) was proven to stimulate natural skin protective functions. The major functions are an increased energy level and the prevention of DNA damage. These functions can I be defined as biological UV protection. The tumor suppressor protein p53 plays a key role in the regulation of DNA repair. p53 must be transferred into the phosphorylated form to work as transcription factor for genes which are regulating the cell cycle or organizing DNA repair. A pretreatment with SSP increases the phosphorylation rate of p53 of chronically UV-irradiated human keratinocytes significantly. According to the same test procedure SSP induces a dramatic increase in the expression of the tumor suppressor protein p14$^{ARF}$ that is supporting the p53 activity by blocking the antagonist of p53, the oncoprotein Mdm2. Mdm2, a ubiquitin E3-ligase, downregulates p53 and at the same time it prevents phosphorylation of p53. The positive influence of the tumor suppressor proteins explains the stimulation of DNA repair and prevention of sunburn cell formation by SSP, which was proven in cell culture experiments. In vivo the increased skin tolerance against UV irradiation by SSP could be confirmed too. We have assumed, that an increased repair potential provides full cell functionality.y.

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Cloning and Characterization of dnaK Heat Shock Protein Gene in a Halotolerant Cyanobacterium (내염성 cyanobacteria로 부터 danK heat shock protein 유전자의 cloning 및 특성 해명)

  • ;;;Teruhiro Takabe
    • Journal of Life Science
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    • v.11 no.5
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    • pp.464-469
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    • 2001
  • A gene, dnaK2, encoding a distinct member of the HSP70 family of molecular chaperones is isolated from the halotolerant cyanobactrium Aphanothece halophytica. The dnak2 gene encodes a molecular wight of 68 kDa polypeptide with predicted 616 amino acid residues. The DnaK2 protein has a structural characteristic of bacterial DnaK homologues and shows high similarity to other HSP70/Dank proteins. The danK2 transcripts are hardly detectable at 28$^{\circ}C$ and strongly induced upon heat stress. It is also found that dnaK2 transcript is increased by high-salinity stress even in the absence of heat stress. These results suggest that the DnaK2 protein plays an important role in protecting A. halophytica against damage caused by salt stress at well as heat stress.

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Confirmation of Drought Tolerance of Ectopically Expressed AtABF3 Gene in Soybean

  • Kim, Hye Jeong;Cho, Hyun Suk;Pak, Jung Hun;Kwon, Tackmin;Lee, Jai-Heon;Kim, Doh-Hoon;Lee, Dong Hee;Kim, Chang-Gi;Chung, Young-Soo
    • Molecules and Cells
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    • v.41 no.5
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    • pp.413-422
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    • 2018
  • Soybean transgenic plants with ectopically expressed AtABF3 were produced by Agrobacterium-mediated transformation and investigated the effects of AtABF3 expression on drought and salt tolerance. Stable Agrobacterium-mediated soybean transformation was carried based on the half-seed method (Paz et al. 2006). The integration of the transgene was confirmed from the genomic DNA of transformed soybean plants using PCR and the copy number of transgene was determined by Southern blotting using leaf samples from $T_2$ seedlings. In addition to genomic integration, the expression of the transgenes was analyzed by RT-PCR and most of the transgenic lines expressed the transgenes introduced. The chosen two transgenic lines (line #2 and #9) for further experiment showed the substantial drought stress tolerance by surviving even at the end of the 20-day of drought treatment. And the positive relationship between the levels of AtABF3 gene expression and drought-tolerance was confirmed by qRT-PCR and drought tolerance test. The stronger drought tolerance of transgenic lines seemed to be resulted from physiological changes. Transgenic lines #2 and #9 showed ion leakage at a significantly lower level (P < 0.01) than ${\underline{n}}on-{\underline{t}}ransgenic$ (NT) control. In addition, the chlorophyll contents of the leaves of transgenic lines were significantly higher (P < 0.01). The results indicated that their enhanced drought tolerance was due to the prevention of cell membrane damage and maintenance of chlorophyll content. Water loss by transpiration also slowly proceeded in transgenic plants. In microscopic observation, higher stomata closure was confirmed in transgenic lines. Especially, line #9 had 56% of completely closed stomata whereas only 16% were completely open. In subsequent salt tolerance test, the apparently enhanced salt tolerance of transgenic lines was measured in ion leakage rate and chlorophyll contents. Finally, the agronomic characteristics of ectopically expressed AtABF3 transgenic plants ($T_2$) compared to NT plants under regular watering (every 4 days) or low rate of watering condition (every 10 days) was investigated. When watered regularly, the plant height of drought-tolerant line (#9) was shorter than NT plants. However, under the drought condition, total seed weight of line #9 was significantly higher than in NT plants (P < 0.01). Moreover, the pods of NT plants showed severe withering, and most of the pods failed to set normal seeds. All the evidences in the study clearly suggested that overexpression of the AtABF3 gene conferred drought and salt tolerance in major crop soybean, especially under the growth condition of low watering.

Overexpression of Ice Recrystallization Inhibition Protein (HvIRIP) from Barley Enhances Cold Tolerance in Transgenic rapeseed plants (HvIRIP 과발현 유채 형질전환체의 내한성 증진)

  • Roh, Kyung Hee;Park, Jong-Sug;Kang, Han-Chul;Kim, Jong-Bum;Jang, Young-Suk;Kim, Kwang-Soo;Yi, Hankuil
    • Journal of Applied Biological Chemistry
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    • v.58 no.4
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    • pp.325-332
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    • 2015
  • Rapeseed (Brassica napus) is now the second largest oilseed crop after soybean. Cold temperature tolerance is an important agronomic trait in winter rapeseed that determines the plant's ability to control below freezing temperatures. To improve cold tolerance of rapeseed plants, an expression vector containing an Barley Ice recrystallization inhibition protein (HvIRIP) cDNA driven by a cauliflower mosaic virus 35S promoter was transferred into rapeseed plants. Transgenic expression of HvIRIP was proved by southern- and northern-blot analyses. The level of freezing tolerance of transgenic $T_3$ plants was found to be significantly greater than that of wild-type rapeseed plants by freezing assay. Proline accumulation during cold stress was also highly induced in the transgenic rapeseed plants. The transgenic plants exhibited considerable tolerance against oxidative damage induced by cold stress. Our results indicated that heterologous HvIRIP expression in transgenic rapeseed plants may induce several oxidative-stress responsive genes to protect from cold stress.