• Environmental Mutagens and Carcinogens
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• v.9 no.1
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• pp.23-32
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• 1989

Chinese hamster ovary (CHO)-K1 cells echibited a differential sensitivity in the process of DNA repair synthesis induced by ethyl methanesulfonate (EMS) or bleomycin (BLM) in relation to cell cycle. Two assays were employed in this study: alkaline elution and unscheduled DNA synthesis. The post-treat-ment with aphidicolin (APC), an inhibitor of DNA polymerase alpha, inhibited DNA repair synthesis induced by EMS in G2 phase, while APC did not show any effect on BLM-induced DNA repair synthesis in all phases. On the other hands, the 2', 3'-dideoxythymidine (ddTTP), an inhibitor of DNA polymerase beta, inhibited DNA repair synthesis induced by EMS or BLM in both of G1 and G2 phases. These results suggested that the involvement of DNA polymerase alpha and beta in DNA repair was dependent on cell stage or used chemical agent.

• Molecular & Cellular Toxicology
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• v.4 no.4
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• pp.285-292
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• 2008

Crocin is one of the major components of gardenia fruit and saffron which are widely used as natural food colorants and as traditional Chinese medicines. However, the genotoxicity data on crocin are not sufficient for safety evaluation. The purpose of this study was the examination of the genotoxicity on crocin from gardenia yellow in bacterial and mammalian cells, using various genotoxic battery testing assays and the influence of crocin on methyl methanesulfonate (MMS) and ${H_2}{O_2}$-induced DNA damage in vitro, using single cell gel electrophoresis (comet) assay. From results, no considerable mutagenicity and clastogenicity were seen in bacteria and mammalian cells treated with crocin, by Ames test, chromosomal aberration assay, ${tk}^{+/-}$ gene forward mutation assay and comet assay. And, post-treatment with crocin significantly suppressed ${H_2}{O_2}$-induced DNA damage in a dose-dependent manner. In conclusion, the findings of the present study and other previous observations indicate that crocin has no genotoxic potential. And it showed that crocin clearly repressed the genotoxic potency of ${H_2}{O_2}$. These results suggest that anti-oxidative effects of crocin may be involved in the protective effects of DNA damage.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.43 no.1
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• pp.28-31
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• 1998

Nodule development was regulated partially by host plant factors originating in the shoots and roots. This study was performed to identify the origin of the factors regulating nodulation in supernodulating soybean (Glycine max [L.] Merr.) mutant 'SS2-2' which was isolated recently from ethyl methanesulfonate (EMS) mutagenesis of 'Sinpaldalkong 2'. Self- and reciprocal-grafts were made among three soybean genotypes which consisted of two supernodulating mutants, SS2-2 and 'nts 382', and a normal nodulating Sinpaldalkong 2. Self-grafted supernodulating mutants were characterized by greater nodule number, nodule dry weight, and $C_2$H$_2$ reduction activity than self-grafted wild types. They were also characterized by relatively higher nodule to root dry weight. Significant shoot genotypic effects were observed on nodule number, nodule dry weight, and $C_2\;H_2$ reduction activity per plant, whereas varying root genotypes had no effects. From this result, it is surmised that supernodulating characters are controlled by a graft-transmissible shoot factor, and mutant SS2-2 may have similar nodulation mechanism to the former supernodulating nts 382. In all grafts, both supernodulating mutants and Sinpaldalkong 2 maintained the similar balance between above ground and below ground parts regardless of significant differences in partitioning of dry matter into root and nodule between supernodulating mutants and Sinpaldalkong 2.

• Molecules and Cells
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• v.22 no.2
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• pp.203-209
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• 2006

TBP (TATA-binding protein)-related factor 2 (TRF2) regulates transcription during a nuber of cellular processes. We previously demonstrated that it is localized in the cytoplasm and is translocated to the nucleus by DNA-damaging agents. However, the cytoplasmic localization of TRF2 is controversial. In this study, we reconfirmed its cytoplasmic localization in various ways and examined its nuclear migration. Stresses such as heat shock, redox agents, heavy metals, and osmotic shock did not affect localization whereas genotoxins such as methyl methanesulfonate (MMS), cisplatin, etoposide, and hydroxyurea caused it to migrate to the nucleus. Adriamycin, mitomycin C and ${\gamma}$-rays had no obvious effect. We determined optimal conditions for the nuclear migration. The proportions of cells with nuclei enriched for TRF2 were 25-60% and 5-10% for stressed cells and control cells, respectively. Nuclear translocation was observed after 1 h, 4 h and 12 h for cisplatin, etoposide and MMS and hydroxyurea, respectively. The association of TRF2 with the chromatin and promoter region of the proliferating cell nuclear antigen (PCNA) gene, a putative target of TRF2, was increased by MMS treatment. Thus TRF2 may be involved in genotoxin-induced transcriptional regulation.

• The Korean Journal of Zoology
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• v.37 no.2
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• pp.232-239
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• 1994

RecA protein plans a central role in homologous recombination and DNA repair in Escherichia cofi (E. colD. The function 8nd structure of this protein are universal in prokarvotes and also conserved in eukaryotes such as yeast. The RecA-like protein with 74 lInDa in size has already been identified and purified from a fission yeast Schizosaccharomyces pombe (5. pommel (Lee, 19911. From this study it was revealed that the RecA-like protein of 5. pombe was highly inducible to various DNA damaging agents and inhibitors of nucleotide pool svnthesizins enzymes. The cellular level of the 5. pombe RecA-like protein wi,u markedly increased, upto 5- to 10-fold, by treatment with various DNA-damains agents including ultraviolet (UV) light, methyl methanesulfonate WS),4-nitroquinoline-1-oxide (4-NQO), and mitomycin-C (MMC), similar to E. cofi RecA protein. Interestingly, the protein level was also increased by inhibitors of nucleotide pool forming enzlwnes such as methotrexate (MTX) and hvdroxvurea (HU). The most effective doses for the inducibility of 4-NQO, MMS, W, MMC, MTX, and HU were 0.2 Ug/ml, 30 mM, 200 J/ma, 0.4 $\mus/ml,$ 1 Ug/ml, and 100 mM, respectively. The range of effective duration time for the inducibilitv of RecA-like protein was from 270 to 450 mins. These results suggest that the 5. pombe RecA-like protein also platys an imortant role in cellular responses to DNA damage as in E. coli system.

• Mycobiology
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• v.39 no.4
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• pp.272-277
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• 2011

Chemical mutagenesis of basidiospores of Hypsizygus marmoreus generated new mushroom strains. The basidospores were treated with methanesulfonate methylester, an alkylating agent, to yield 400 mutant monokaryotic mycelia. Twenty fast-growing mycelia were selected and mated each other by hyphal fusion. Fifty out of the 190 matings were successful (mating rate of 26.3%), judged by the formation of clamp connections. The mutant dikaryons were cultivated to investigate their morphological and cultivation characteristics. Mutant strains No. 3 and No. 5 showed 10% and 6% increase in fruiting body production, respectively. Eight mutant strains showed delayed and reduced primordia formation, resulting in the reduced production yield with prolonged cultivation period. The number of the fruiting bodies of mutant No. 31, which displayed reduced primordial formation, was only 15, compared to the parental number of 65. Another interesting phenotype was a fruiting body with a flattened stipe and pileus. Dikaryons generated by mating with the mutant spore No. 14 produced flat fruiting bodies. Further molecular biological studies will provide details of the mechanism. This work shows that the chemical mutagenesis approach is highly utilizable in the development of mushroom strains as well as in the generation of resources for molecular genetic studies.

• Animal cells and systems
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• v.4 no.3
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• pp.293-297
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• 2000

The present investigation was performed to elucidate the effect of pretreatment with low dose ultraviolet radiation (UV) and ethyl methanesulfonate (EMS) on cell survival by trypan blue dye exclusion method and plasma membrane glycoconjugates by lectin-cytochemistry in sarcoma 180 (S180) cells. Pretreatment with 2 J/$m^2$ UV or 2 mM EMS increased the percentage of survival of cells subsequently treated with high dose UV (10 or 20 J/$m^2$) or EMS (10 or 20 mM), respectively. Staining intensity of concanavalin A (Con A) of the cells pretreated with 2J/$m^2$ UV or 2 mM EMS and subsequently treated with 10 or 20 mM EMS was stronger than that of the cellstreated with 10 or 20 mM EMS. These results suggest that there is an adaptive response on cell survival to EMS or UV in S180 cells. And the results show a change in mannose-containing glycoconjugates of plasma membrane in S180 cells pretreated with EMS or UV and subsequently treated with EMS.

• The Korean Journal of Zoology
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• v.19 no.2
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• pp.71-78
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• 1976

Chromosome aberration induced by methyl methanesulfonate (MMS) and the effects of thymidine analogs (BUdR or IUdR) on MMS-induced chromosome aberration were studied in human lymphocyte cultures. Single treatment with MMS to lymphocytes induces both chromatid and chromosome type aberrations with high frequency of chromatid type. The combined treatment of BUdR or IUdR with MMS was found to be more effective in increasing the rate of chromosome type aberrations.

• Development and Reproduction
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• v.23 no.2
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• pp.183-191
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• 2019

The objective of this study is to evaluate the anesthetic effects of clove oil and tricaine methanesulfonate (MS-222) on the Far Eastern catfish, Silurus asotus, by measuring the times to anesthesia and recovery. Each anesthetic effect of clove oil and MS-222 was tested in two groups of fish with different body sizes: a group of small fish (mean body length: $15.5{\pm}1.58cm$, mean body weight: $50.1{\pm}5.91g$, n=20) and a group of large fish (mean body length: $31.5{\pm}4.19cm$, mean body weight: $302.1{\pm}15.22g$, n=20). The anesthetics were used at concentrations of 200, 300, 400, 500, and 600 ppm. The results showed significant relationships between the concentration of the anesthetic and the body size of the fish. Each of these variables showed statistical significance (p<0.05). The time to anesthesia decreased linearly with increasing concentration in the large fish for both clove oil and MS-222 (p<0.05). Based on an optimal anesthetic time of approximately 1 min, the preferred concentrations of the anesthetics were 500 ppm for clove oil and 600 ppm for MS-222. Both the anesthetic time and the recovery time were shorter for the small fish than for the large fish (p<0.05). Our study showed that the smaller-sized Far Eastern catfish was more easily anesthetized and recovered more rapidly from anesthesia than the larger-sized fish.

• Animal cells and systems
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• v.3 no.2
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• pp.229-236
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• 1999

The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.