• Journal of Nutrition and Health
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• 제32권6호
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• pp.654-660
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• 1999

Folate, a precursor of the coenzyme tetrahydrofolate, plays an important role in DNA replication and cell proliferation, and thus could influence rapidly proliferating immune cells such as leukocytes and splenocytes. The effects of dietary folate on folate concentrations of plasma, thymus, spleen and leukocytes were investigated in rats. The animals were raised for 6 weeks on semipurifed experimental diets containing 0mg, 2mg, 4mg, 8mg folate/kg diet. Folate concentrations were determined microbiologically using Lactobacillus casei(ATCC 7469), and DNA strand breaks produced by alkaline treatment were analyzed fluorometrically. When compared to folate adequate diet, the folate deficient diet(0mg folate/kg diet) resulted in lowest folate levels in plasma, thymus, spleen and leukocytes, and the highest DNA strand breaks in spleen cells and leukocytes. Dietary folate levels significantly increased folate concentrations of immune tissues, leukocytes, and the plasma in a dose dependent manner, folate concentrations being highest with a diet providing 8mg folate/kg diet. The percentages of the double strand DNA remaining in the splenocytes and leukocytes after alkaline treatment were significantly increased with higher amounts of dietary folate in a dose dependent manner. Folate intakes of 8mg than 4mg/kg diet was found to be more effective in the prevention of DNA strand breaks. The results of this study suggest that increased folate above the requirement level could improve DNA stabilities in immune cells.

• Animal cells and systems
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• 제6권4호
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• pp.311-315
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• 2002

The RAD4 gene is essential for nucleotide excision repair in Saccharomyces cerevisiae. It has been known that the deduced amino acid sequence of Rad4 protein contains three DNA-dependent ATPase/helicase motifs. To determine the biochemical activities and functional role of RAD4 the Rad4 protein was expressed and purified. Immunoblot analysis showed a specific band of 21 kDa, which was well-matched with the size of open reading frame of the RAD4 gene. The purified Rad4 protein had no detectable helicase activity. However, the protein could interact with double stranded oligonucleotides, as judged by mobility shift assay. This result suggests that the Rad4 protein is a DNA binding protein.

• Journal of Radiation Protection and Research
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• 제20권2호
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• pp.71-78
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• 1995

본 연구에서는 유전자 손상회복에 관여하는 단백질을 이용하여 돌연변이 생성을 억제시키는 물질로서 알려진 cobaltous chloride가 유전자 손상회복에 미치는 영향을 연구하므로서 방사선으로 인한 손상방지 및 방사선 방어효과에 대한 적용가능성을 평가하였다. Cobaltous chloride가 RecA 단백질의 기능에 미치는 영향을 조사한 결과 RecA 단백질에 의한 DNA strand exchange 반웅에 있어 cobaltous chloride 처리로 RecA 단백질이 $_{ss}DNA$로 부터 SSB 단백질과 더 효과적으로 경쟁함으로써 안정된 $RecA-_{ss}DNA$ complex의 형성을 유도하고, 증가된 ATPase활성에 의한 ATP 가수분해로 손상된 DNA의 회복이 촉진될 수 있다는 사실을 입증 해주고 있다. 또한 RecA단백질은 UV에 의해 손상된 supercoiled DNA에 더 효과적으로 결합됨이 관찰되었으며 UV 선량과도 상관관계가 있음을 확인하였다. 따라서 이와 같은 연구결과들은 방사선으로 인한 유전적인 손상방지 및 방사선 방어효과에 관한 연구에 적용될 수 있을 것으로 기대된다.

• Archives of Pharmacal Research
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• 제27권1호
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• pp.77-82
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• 2004

DA-125, a novel derivative of adriamycin, is known for its anti-cancer activity. In this study, the inhibitory mechanism of DA-125 on topoisomerase was investigated in the simian virus 40 (SV40) replicating CV-1 cell by studying the SV40 DNA replication intermediates and DNA-topoisomerase complexes. DNA-protein complexes that were formed in the drug-treated cells were quantitated by using a glass filter assay. SV40 DNA replication intermediates that were accumulated in the drug-treated CV-1 cell were analyzed in a high resolution gel. DA-125 did not accumulate B-dimers of SV40 DNA replication intermediates which were found in the adriamycin-treated CV-1 cells. DA-125 induced a dose-dependent formation of the DNA-protein complexes, while adriamycin did not. When adriamycin and etoposide (VP16) were added to the SV40-infected cells at the same time, adriamycin blocked the formation of the DNA-protein complexes induced by VP16 in a dose-dependent manner. However, DA-125 blocked the formation of the DNA-protein complexes induced by VP16 up to the maximum level of the DNA-protein complexes that were induced by DA-125 alone. Adriamycin and DA-125 did not inhibit the formation of the DNA-protein complexes that were caused by camptothecin, a known topoisomerase I poison. DA-125 is bifunctional in inhibiting topoisomerase II because it simultaneously has the properties of the topoisomerase II poison and the DNA intercalator. As a topoisomerase II poison, DA-125 alone induced dose-dependent formation of the DNA-protein complexes. However, as a DNA intercalator, it quantitatively inhibited the formation of the DNA-protein complexes induced by a strong topoisomerase II poison VP16. Furthermore considering that the levels of the DNA-protein complex induced by VP16 were decreased by DA-125 in terms of the topoisomerase II poison, we suggest that DA-125 has a higher affinity to the drug-binding sites of DNA than VP16 has.

• 미생물학회지
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• 제43권4호
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• pp.285-291
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• 2007

볼티모어의 분류체계에 의하면 바이러스는 복제 및 단백질합성 전략에 따라 6개의 집단으로나눌 수 있다. 몇 종류의 작은 DNA 바이러스를 제외한 대부분의 바이러스는 게놈 복제를 위한 자신의 핵산중합효소를 유전자로 암호화하고 있다. 바이러스 핵산중합효소에는 DNA-의존DNA 중합효수, RNA-의존RNA 중합효소, RNA-의존 DNA 중합효소 세 종류가 있으며, 이들은 모두 4개의 공통된 모티프(motif)를 가진다. 우리는 볼티모어의 분류체계와 바이러스의 핵산중합효소와의 관계를 아미노산 서열을 통해 분자 계통분류학적 분석을 통해 알아보고자 하였다. NCBI GenBank에서 얻은 바이러스 중합효소의 아미노산 서열을 CLUSTAL X 프로그램으로 다중서열하고, Neighbor-joining, Maximum-likelihood, Bayesian의 세 가지 방법으로 계통도를 그려보았다. 미세한 차이는 있었으나, 세 가지 방법 모두에서 볼티모어의 분류법과 일치하는 결과를 보였고, 특이하게도 두 가닥 RNA 바이러스는 숙주의 종류에 따라, (-)RNA 바이러스는 게놈의 절편화에 따라 각각2개의 소집단으로 나뉘어지는 것을 볼 수 있었다.

• Toxicological Research
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• 제14권4호
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• pp.525-533
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• 1998

The covalent interactions of toluenediisocyanate (TDI) with macromolecules were investigated both in vitro and in vivo. In vitro incubations of 2,4- and 2,6-TDI with DNA or proteins resulted in dose-dependent formation of TDI-protein and TDI-DNA adducts. TDI-treated DNA was highly resistant to enzymatic digestion and thermal hydrolysis, but was readily hydrolyzed under acidic conditions by releasing its corresponding toluenediamine (TDA), suggesting that TDI caused the crosslinking of DNA. Reaction of TDI with albumin and globin resulted in the formation of several adducts, and some adducts were formed in blood of TDI-treated rats in a dose-dependent fashion. Administration of TDI to rats resulted also in a dose-dependent binding of TDI to hepatic tissue. Levels of TDI-albumin adducts were 10 times higher than those of TDI-globin adducts; the biological half lives of TDI-albumin and TDI-globin adducts were 1.2 and 12.5 days, respectively. Globin adducts were detected up to 28 days after the treatment. Hepatic TDI protein adducts were persistent for a substantial period whereas the levels of hepatic TDI-DNA adduct were decreased rapidly. These results indicate that the isocyanato group of TDI is not readily hydrolyzed under physiological conditions, is transported to other organs, and is bound to DNA and/or proteins without further metabolic activation. As the adducted products degrade in the body, TDA is released and introduced to the liver. TDA may additionally bind to hepatic tissue after metabolic activation. Thus, the toxic effect of TDI exposure is considered to persist during the lifetime of the adducted biological macromolecules.

• 한국동물학회지
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• 제15권3호
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• pp.95-100
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• 1972

回復複製(unscheduled DNA synthesis or repair replication)와 染色體交換 및 分裂活動과의 相關關係를 추구하기 위해 X-線을 處理한 4種의 哺乳動物 細胞를 材料로 回復複製 時期와 線量反應을 調査하였다. 回復複製는 調査한 4種의 細胞에서 모두 일어나고 그 率은 照射線量에 比例하며 細胞 種類에 따라 差가 난다. 그러나 回復複製가 일어나는 時期는 細胞 種類에 관계없이 X-線 照射후 2時間까지 지속되고 있다. 또 回復複製의 相對量은 細胞의 種族染色體數, 分裂活動 및 染色體交換率과 직접적인 관련이 없다. 分裂活動能力과 染色體交換은 다 線量에 比例하나 그 率은 逆比例 관계를 나타낸다.

• 대한화장품학회지
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• 제48권1호
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• pp.33-38
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• 2022

자외선인 ultraviolet B (UVB)는 피부각질세포의 DNA 잔기에 손상을 준다. 특히, DNA의 pyrimidine 잔기 손상인 cyclobutane pyrimidine dimers (CPD)의 형성은 피부 광노화의 대표적인 지표로 여겨진다. 본 연구에서는 피부 각질세포에서 UVB에 의한 DNA 손상을 완화 시키는 소재로 감초추출물(Glycyrrhiza glabra extract, G. glabra extract)의 효능을 확인하였다. 먼저 피부각질세포에서 UVB 의존적으로 CPD형성이 증가하는 것을 확인하였다. 이후 감초추출물에 의해 UVB 유발 CPD 형성이 유의하게 줄어드는 것을 확인할 수 있었다. 추가로 DNA 손상회복 인자의 mRNA 발현이 감초추출물에 의해 증가하는 것도 확인하였다. 결론적으로 본 연구를 통해 감초추출물의 피부각질세포에서의 DNA 보호 효과를 확인할 수 있었다.

• 대한치과교정학회지
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• 제21권1호
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• pp.97-111
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• 1991

It is the aim of this study to investigate the effects of sodium fluoride and sodium orthovanadate upon the proliferation and activity of the osteoblast (MC3T3-E1 cells). MC3T3-E1 cells were cultured in $\alpha-MEM$ containing $10\%$ FBS and various concentration of sodium fluoride and sodium orthovanadate was appended to serum free media. DNA synthesis was examined through the $[^3H]$ thymidine incorporation into DNA. Collagen synthesis was examined through the $[^3H]$ proline incorporation into collagenase digestible protein and noncollagen protein. The following results were drawn; 1. Sodium fluoride stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M$ to $10{\mu}M$ (P < 0.005). 2. Sodium orthovanadate stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M\;to\;8{\mu}M$, however showed diminution at $10{\mu}M$ (P < 0.001). 3. Sodium fluoride and sodium orthovanadate stimulated the percent collagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M$ to $10{\mu}M$ (P < 0.001). 4. Sodium fluoride and sodium orthovanadate stimulated the noncollagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M\;to\;10{\mu}M$ (P < 0.001). In conclusion, sodium fluoride and sodium orthovanadate stimulate the proliferation and activity of osteoblast by stimulation of DNA synthesis and collagen and noncollagen synthesis in osteoblast.

• Molecules and Cells
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• 제39권9호
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• pp.705-713
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• 2016

The efficiency of Agrobacterium-mediated transformation in plants depends on the virulence of Agrobacterium strains, the plant tissue culture conditions, and the susceptibility of host plants. Understanding the molecular interactions between Agrobacterium and host plant cells is crucial when manipulating the susceptibility of recalcitrant crop plants and protecting orchard trees from crown gall disease. It was discovered that Arabidopsis voltage-dependent anion channel 1 (atvdac1) mutant has drastic effects on Agrobacterium-mediated tumorigenesis and growth developmental phenotypes, and that these effects are dependent on a Ws-0 genetic background. Genetic complementation of Arabidopsis vdac1 mutants and yeast porin1-deficient strain with members of the AtVDAC gene family revealed that AtVDAC1 is required for Agrobacterium-mediated transformation, and there is weak functional redundancy between AtVDAC1 and AtVDAC3, which is independent of porin activity. Furthermore, atvdac1 mutants were deficient in transient and stable transformation by Agrobacterium, suggesting that AtVDAC1 is involved in the early stages of Agrobacterium infection prior to transferred-DNA (T-DNA) integration. Transgenic plants overexpressing AtVDAC1 not only complemented the phenotypes of the atvdac1 mutant, but also showed high efficiency of transient T-DNA gene expression; however, the efficiency of stable transformation was not affected. Moreover, the effect of phytohormone treatment on competence to Agrobacterium was compromised in atvdac1 mutants. These data indicate that AtVDAC1 regulates the competence of Arabidopsis to Agrobacterium infection.