• Journal of Ginseng Research
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• v.44 no.5
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• pp.747-755
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• 2020

Background: Ginsenosides accumulation responses to temperature are critical to quality formation in cold-dependent American ginseng. However, the studies on cold requirement mechanism relevant to ginsenosides have been limited in this species. Methods: Two experiments were carried out: one was a multivariate linear regression analysis between the ginsenosides accumulation and the environmental conditions of American ginseng from different sites of China and the other was a synchronous determination of ginsenosides accumulation, overall DNA methylation, and relative gene expression in different tissues during different developmental stages of American ginseng after experiencing different cold exposure duration treatments. Results: Results showed that the variation of the contents as well as the yields of total and individual ginsenosides Rg1, Re, and Rb1 in the roots were closely associated with environmental temperature conditions which implied that the cold environment plays a decisive role in the ginsenoside accumulation of American ginseng. Further results showed that there is a cyclically reversible dynamism between methylation and demethylation of DNA in the perennial American ginseng in response to temperature seasonality. And sufficient cold exposure duration in winter caused sufficient DNA demethylation in tender leaves in early spring and then accompanied the high expression of flowering gene PqFT in flowering stages and ginsenosides biosynthesis gene PqDDS in green berry stages successively, and finally, maximum ginsenosides accumulation occurred in the roots of American ginseng. Conclusion: We, therefore, hypothesized that cold-induced DNA methylation changes might regulate relative gene expression involving both plant development and plant secondary metabolites in such cold-dependent perennial plant species.

• Journal of Life Science
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• v.11 no.2
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• pp.111-115
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• 2001

This study was designed to clone the SNF2/SW12 helicase-related genes from the fission yeast Schizosaccha-romyces pombe and thereafter to elucidate the common functions of the proteins in this family. The $hrp^{2+}$gene was cloned by polymerase chain reaction amplification using degenerative primers from conserved SNF2 motifs within the ERCC6 gene, which encodes a protein involved in DNA excision repair. Like other SNF2/SW12 family proteins, the deduced amino acid sequence of Hrp2 contains DNA-dependent ATPase/7 helicase domains as well as the chromodomain and the DNA binding domain. This configuration is similar to that of mCHD1 (mouse chromo-ATPase/helicase-DNA-dinding protein 1), suggesting that Hrp2 is a S. pombe homolog of mCHD1, which is thought to function in altering the chromatin structure to control the gene expression. To characterize the function of Hrp2, 4 Uracil-Hrp2 fusion protein, it was purified near homogeneity by affinity chromatography on $Ni^{2+}$-NTA agarose, DEAE-Sepharose ion exchange arid Sephacryl S-200 gel filtration chromatographies. The purified fusion protein exhibited DNA-dependent ATPase activity, which was stimulated by both double-stranded and single-stranded DNA. To determine the steady-state level of $hrp^{2+}$ transcripts during growth, cells were cultured in medium and collected at every 2hr to prepare total RNAs. The northern blot analysis showed that the level of $hrp^{2+}$ transcripts reached its maximum before the cells entered the exponential growth phase and then decreased gradually, This result implies that Hrp2 may be required at early stages of cell growth.h.

• Journal of Applied Biological Chemistry
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• v.43 no.3
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• pp.156-160
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• 2000

The possible mechanism of the DNA strand breaking activity of the hot-water extract of Fructus Chaenomelis (dried fruit of Chaenomeles sinensis) in a closed circular duplex replica form DNA (RFI DNA) was studied through agarose gel electrophresis under various conditions. Induction of DNA strand scission by the hot-water extract of C. sinensis occurred in dose and time-dependent manners. $Cu^{2+}$ was indispensable for the induction of DNA strand breakage. Exogeneous chelating agents inhibited the DNA breaking activity, conforming the catalytic action of $Cu^{2+}$ on generation of free radicals responsible for oxidative damage. Antioxidant enzymes and some radical scavengers were used to investigate the major radical species triggering the DNA strand scission, demonstrating that a highest inhibitory activity was found in the presence of catalase, while less in the presence of tiron (a scavenger for superoxide radical), 2-aminoethyl-isothiuroniumbromide-HBr, cysteamine (scavengers for hydroxyl radical), and 1,4-diazabicyclo [2,2,2] octane (a scavenger for singlet oxygen) in decreasing order. The findings implied that oxygen radical species generated in presence of transition divalent cation during the oxidation of some compounds contained in the hot-water extract of C. sinensis is mainly responsible for inducing genotoxicity.

• Animal cells and systems
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• v.7 no.2
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• pp.159-164
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• 2003

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. The RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA and DNA-RNA helicase activities. To examine the extent of conservation of structure and function of a S. pombe RAD3 during eukaryotic evolution, the RAD3 homolog gene was isolated by screening of genomic DNA library. The isolated gene was designated as HRD3 (homolog of RAD3 gene). Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as HRD3 gene and this gene exists as a single copy in S. pombe. The transcript of 2.8 kb was detected by Northern blot analysis, The level of transcripts increased by ultraviolet (UV) irradiation, indicating that HRD3 is one of the UV-inducible genes in S. pombe. Furthermore, the predicted partial sequence of HRD3 protein has 60％ identity to S. cerevisiae RAD3 gene. This homology was particularly striking in the regions identified as being conserved in a group of DNA helicases. Gene deletion experiments indicate that the HRD3 gene is essential for viability and DNA repair function. These observations suggest evolutionary conservation of other protein components with which HRD3 might interact in mediating its DNA repair and viability functions.

• Reproductive and Developmental Biology
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• v.32 no.1
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• pp.33-38
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• 2008

DNA methylation is involved in tissue-specific gene control and essential for normal embryo development Octamer-binding transcription factor 4 (Oct-4) is one of the most important transcription factors for early differentiation. This study was performed whether the bovine Oct-4 is tissue specific or developmental dependent epigenetic mark, we investigated transcripts and the methylation status of CpGs of 5'-promoter region of Oct-4 in bovine preimplantation embryos. Oct-4 transcripts were highly detected in morula and blastocyst, while they were present low levels in sperm and 2- to 8-cell stage embryos. These results suggest that de novo expression of Oct-4 initiates at morula stage of embryogenesis. Here we determined that there is a tissue-dependent differentially methylated region (T-DMR) in the 5'-promoter region of Oct-4. The methylation status of the Oct-4 T-DMR was distinctively different in the oocyte from that in the sperm and adult somatic tissues and changed from zygote to blastocyst stage, suggesting that active methylation and demethylation occur during preimplantation development. Based on these results, the 5'-promoter region of Oct-4 gene is target for DNA methylation and the methylation status changes variously during embryonic development in bovine.

• Journal of Life Science
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• v.15 no.4 s.71
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• pp.542-547
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• 2005

Hypersensitivity of cells lacking Brcal to DNA interstrand .ross-link (ICL) agents such as cisplatin and mitomycin C(MMC) implicates the important role of Brcal in cellular response following ICL treatment. Brca1 plays an essential role in DNA double-strand break (DSB) repair through homologous recombination (HR)-dependent and -independent process. Recently, our group has been reported that Brca1 involves in cellular ICL response through HR-dependent repair process (Yun J. et at., Oncogene 2005). In this report, the involvement of Brca1 protein in HR-independent repair process is examined using isogenic $p53^{-/-}\;and\;p53^{-/-}\;Brcal^{-/-}$ mouse embryonic fibroblast (MEF) and psoralen cross-linked reporter reactivation assay. Brcal-deficient MEFs showed significantly low HR-independent repair activity compare to Brca1-proficient MEFs. Hypersensitivity to MMC and ICL reporter repair activity were restored by the reconstitution of Brca1 expression. Interestingly, MEFs expressing exon 11-deleted isoform of Brca1 $(Brca1^{\Delta11/\Delta11})$ showed high resistance to MMC and ICL reporter repair activity comparable to Brca1-reconstituted MEFs. Taken together, these results suggest that Brca1 involves in ICL repair through not only HR-dependent process but also HR-independent process using N-terminal RINC finger domain or C-terminal BRCT domain rather than exon 11 region which mediate interaction with Rad50.

• Proceedings of the Korean Institute of Intelligent Systems Conference
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• 2000.11a
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• pp.107-110
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• 2000

In this paper, we propose a method of constructing neural networks using bio-inspired emergent and evolutionary concepts. This method is algorithm that is based on the characteristics of the biological DNA and growth of plants. Here is, we propose a constructing method to make a DNA coding method for production rule of L-system. L-system is based on so-called the parallel rewriting mechanism. The DNA coding method has no limitation in expressing the production rule of L-system. Evolutionary algorithms motivated by Darwinian natural selection are population based searching methods and the high performance of which is highly dependent on the representation of solution space. In order to verify the effectiveness of our scheme, we apply it to one step ahead prediction of Mackey-Glass time series.

• Toxicological Research
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• v.20 no.1
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• pp.63-69
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• 2004

Casapse-3 protease is known as a key role of apoptotic enzyme, and caspase-3 activity is a central event that occurs upstream of DNA fragmentation during apoptosis. This study demonstrates that chitosan pretreatment inhibits cadmium-induced apoptosis by attenuating the activity of caspase-3. We also analyzed the protective effect of chitosan on DNA fragmentation induced by cadmium. Cadmium toxicity was examined by DNA fragmentation and nuclear condensation with Hoechst stain. Caspase-3 activities were increased cadmium treated group for 3 hours compared with control. When chitosan (150 mg/ml) was pretreated at 30 min before cadmium treatment, cadmium cytotoxicity was suppressed in a dose-dependent manner evaluated by DNA fragmentation and caspase activity. From these results, it is suggest that the protective effect of chitosan pretreatment against cadmium-induced cytotoxicity is mediated through inhibition of caspase-3 protease activation and DNA fragmentation.

• Proceedings of the Korean Institute of Intelligent Systems Conference
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• 2000.05a
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• pp.224-227
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• 2000

In this Paper, we prepose a method of constructing neural networks using bio-inspired emergent and evolutionary concepts. This method is algorithm that is based on the characteristics of the biological DNA and growth of plants. Here is, we propose a constructing method to make a DNA coding method for production rule of L-system. L-system is based on so-called the parallel rewriting mechanism. The DNA coding method has no limitation in expressing the production rule of L-system. Evolutionary algorithms motivated by Darwinian natural selection are population based searching methods and the high performance of which is highly dependent on the representation of solution space. In order to verify the effectiveness of our scheme, we apply it to one step ahead prediction of Mackey-Glass time series, Sun spot data and KOSPI data.

• Journal of the Korean Institute of Intelligent Systems
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• v.10 no.4
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• pp.315-323
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• 2000

In this paper, we propose a method of constructing neural networks using bio-inpired emergent and evolutionary concepts. This method is algorithm that is based on the characteristics of the biological DNA and growth of plants, Here is, we propose a constructing method to make a DNA coding method for production rule of L-system. L-system is based on so-called the parallel rewriting nechanism. The DNA coding method has no limitation in expressing the produlation the rule of L-system. Evolutionary algotithms motivated by Darwinaian natural selection are population based searching methods and the high performance of which is highly dependent on the representation of solution space. In order to verify the effectiveness of our scheme, we apply it one step ahead prediction of Mackey-Glass time series, Sunspot data and KOSPI data.