• 제목/요약/키워드: DNA incorporation

검색결과 166건 처리시간 0.021초

생쥐 배아의 전사와 발생에서 DNA/RNA 메틸화의 역할

  • 김종월
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 1998년도 제4차 학술발표대회 및 정기총회
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    • pp.32-33
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    • 1998
  • 생물체에서 유전외적 변형의 하나인 DNA 메틸화는 cis-acting factor의 조성변화를 통하여 세포특이 유전자의 발현과 virus latency, genomic imprinting, mutagenesis등과 같은 생물학적 효과를 나타내는 것으로 알려져 있다.(reviewed by Olle Heby, 1995). 5-azaCR, 5-azaCdR 그리고 6-azaCR의 처리결과는 배아자체의 DNA 메틸화의 유지가 정상발생에 필수적임을 알 수 있으며, 메틸화에 의한 배아발생 조절기작이 존재함을 암시하고 있다. 이러한 과정에서 5-azaCR과 5-azaCdR은 서로다른 경로를 통하여 배아발생에 관여함을 보여주었다. 즉, 5-azaCdR은 주로 DNA에 incorporation되어 작용하는 것으로 여겨지며, 5-azaCR은 DNA 보다는 RNA에 incorporation되어 작용하는 것으로 나타났다. 그리고, 비록 소수의 유전자만이 조사되었지만, 5-azaCdR의 incorporation에 의한 cis-acting factor의 변화는 전사인자인 c-myc proto-oncogene과 fluid 수송에 관여하는 $Na^{+}$, $K^{+}$-ATPase 유전자의 전사를 억제하지 않았다. 반면, 5-azaCR의 RNA로의 incorporation은 전사인자인 c-myc proto-oncogene의 전사를 억제하였으며, 연이어 fluid 수송에 관련되어있는 $Na^{+}$, $K^{+}$-ATPase 유전자의 전사를 억제하였다. 이것은 아마도 RNA로 incorporation된 5-azaCR은 RNA의 post-transcriptional processing에 영향을 주어 trans-acting factor의 조성을 변화, 전사적 repression을 유발한 것으로 사료된다. 생쥐 착상전 초기배아에서 DNA 메틸화는 short-term하게는 cis-acting factor로써 전사적 수준에서 유전자발현 조절하며, 그리고 유전자발현을 통하여 long-term하게는 배아발생에 관여 할 것이라고 사료된다.

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인삼에서 추출한 글리코시드가 배양한 마우스 섬유아세포의 핵산합성에 미치는 영향 (Antiproliferative Effects of Panu ginseng Glycosides on DNA Synthesis in Cultured Mouse Fibroblasts)

  • Byun, Boo-Hyeong;Shin, In-Cheol;Lee, Lil-Ha;Joe, Cheol-O
    • Journal of Ginseng Research
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    • 제19권2호
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    • pp.114-116
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    • 1995
  • Panax ginseng ginsenosides were examined for their affects on the DNA synthesis. The DNA 1 synthesis was measured by the thymidine incorporation into NIH3T3 cells. The ginsenoside, panaxytriol, $Rh_1$ and $Rh_2$ showed reduced [$^{3}H$]-thymidine incorporation. However, other ginsenosides of $Rh_1$, $Rh_2$ and $Rh_3$ did not inhibit DNA synthesis. Among the various ginsenosides, ginsenoside $Rh_2$ was found to be the most inhibitory on DNA synthesis. We suggest $Rh_2$ as one of the potential choice of antiproliferative drugs.

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放射線을 照射한 흰쥐의 여러 가지 組織내의 DNA의 合成과 分解 (Postirradiation Synthesis and Degradation of DNA in Various Tissues of Rats)

  • Kang, Man-Sik
    • 한국동물학회지
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    • 제14권4호
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    • pp.199-204
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    • 1971
  • 放射線照射에 의해서 尿 및 血液內에 다량의 deoxycytidine (CdR)이 流出하는 것을 볼수가 있는데, 이것이 DNA의 合成이나 分解와 어떤 관련성이 있으며, 아울러 어느 臟器의 DNA가 放射線感受性이 큰지의 여부를 밝히기 위해서 400R의 X 線을 흰쥐에 全身照射시켜, 간, 지라 및 흉선을 切除하여 均質化시킨 후, CdR-2-$^14 C$을 써서 DNA의 合成率 및 分解率을 測定하였다. DNA의 合成率은 흉선의 경우, 照射후 1 $\\sim$ 3日에 가장 심한 抑制現象이 나타났으며 5日후 부터는 점차 回復됨을 볼 수 있었고, 한편 간과 지라의 경우는 抑制의 정도가 흉선에 비해서 적었으며 回復도 훨씬 빨리 이루어졌다. DNA의 分解率은 지라와 흉선의 경우 비슷하여서 照射후 1日에 영향이 極大로 나타냈으며 回復도 아주 늦게 일어남을 볼 수 있었는데 반해서, 간의 경우는 分解도 덜 일어났으며 回復도 아주 빨리 일어남을 알 수 있었다. 이와 같은 결과는 각종 臟器의 放射線感受性의 차와 再生能의 차에 의해서 나타나는 현상임을 알 수 있고 CdR의 流出量의 增加는 放射線에 의한 DNA合成의 抑制와 DNA分解의 促進의 두가지 요인에 기인하는 것으로 추정되었으며, 전자의 경우는 흉선이, 후자의 경우는 지라와 흉선이 같은 정도의 重要性을 지니고 있음을 알 수 있었다.

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Biochemical Characterization of the Herpes Simplex Virus-1 DNA Polymerase

  • Song, Byeong-Doo;Lehman, I. Robert
    • BMB Reports
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    • 제32권5호
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    • pp.492-496
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    • 1999
  • We have investigated the biochemical properties of the herpes simplex virus type 1 (HSV-1) DNA polymerase without the UL42 protein (Pol), purified from insect cells infected with a recombinant baculovirus containing the UL30 gene. BSA and DTT have inhibitory effects on dAMP incorporation. Pol showed a greater turnover rate of steady-state single nucleotide incorporation at 12 mM $MgCl_2$ than at 2 mM $MgCl_2$. However, it showed a greater processivity of DNA synthesis at lower $MgCl_2$ concentration (1 mM, 2 mM) than at a higher $MgCl_2$ concentration (12.5 mM). These results are consistent with a slow DNA dissociation at lower $MgCl_2$ concentrations. Pol does not incorporate a correct nucleotide into the primer with an incorrect nucleotide at the end; instead, it preferentially excises the incorrect nucleotide at the 3' end of the primer. Pol has DNA polymerase activity at pHs 6.5 and 7.5 but little at pHs 5.5, 8.5, and 9.5. It has exonuclease activity at pHs 6.5, 7.5, and 8.5 but little at pHs 4.5, 5.5, and 9.5. The finding that Pol has exonuclease activity but not DNA polymerase at pH 8.5 suggests that DNA binds to Pol, but deoxynucleotide binding or incorporation does not occur at pH 8.5.

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Inhibitory Effect of Kale Juice on the Growth and DNA Incorporation of Human Cancer Cells

  • Lee, Seon-Mi;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • 제2권2호
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    • pp.167-173
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    • 1997
  • The inhibitory effects of kale juice on the growh and DNA incorporation of human cancer cells, using HT-29 colon cancer cells, MG-63 osteosarcoma cells, AGS gastric adenocarcinoma cells and K-562 leukemia cells, were studied. The growth of human cancer cells were inhibited in the presence of kale juice (10, 20 nd 40$\mu$l/ml) and the effects were the juice concentration- and incubation time-dependent up to 6 days. When 20$\mu$l/ml of kale juice was added to the media of HT-29, MG-63, AGS and K-562 cancer cells, the cell growth after 6 or 4 days of incubation was retarded by 83~95% of control group. Morphological changes of HT-29 colon cancer cells wre studied under inverted microscope. As the concentration of kale juice increased up to 20$\mu$l/ml, degree of cell aggregation was decreased. Moreover, the DNA incorporation o AGS gastric adenocarcinoma cells and MG-63 osteosarcoma cells which were labeled with [$^3$H] thymidine was significantly reduced after 2 days of incubation at 37$^{\circ}C$ with kale juice. Therefore, we concluded that kale juice strongly decreased the growth of various human cancer cells.

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흰쥐의 소장 점막 세포의 성장에 미치는 Glutamine, Glycine과 Nucleosides/Nucleotide 혼합물의 효과 (Effects of Glutamine, Glycine and Nucleosides/Nucleotide Mixture on Intestinal Mucosal Growth in Rats)

  • 이선영;오현인
    • 한국식품영양과학회지
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    • 제26권1호
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    • pp.130-136
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    • 1997
  • 250g내외의 횐쥐에게 Thiry-Vella fistula를 만든 후 수술과 비경구 영양 효과로 인하여 위축되는 소장의 누공에 각각 nucleosides/nucleotide혼합용액 (Nuc군)이나 glutamine(Gln군), glycine(Gly군)등을 투여하여 소장 점막의 위축 방지나 재생효과가 있는지를 알아보고 자 실험하여 다음과 같은 결과를 얻었다. Thiry-Vella fistula 시술 후 9일 동안 2일에 한 번씩 총 4회 소장 누공으로 주입한 이미노산들과 nucleosides/nucleotide 혼합물의 효과는 누공의 소장 점막에 미치는 직접적인 효과와 비누공 부분에 대한 간접적인 효과로 나누어 볼 수 있다. Nucleosides/nucleotide 혼합물은 소장 누공으로 주입되어 직접 접촉되는 부분의 소장 DNA함량을 유의적으로 증가 시켰으며 단백질 함량도 높은 수치를 보여 주었고 조직 검사 결과 소장 융모의 길이와 표면적을 증가시킨 것으로 나타났다. 그러나 [$^3$H] thymidine incorporation의 결과는 대조군과 유의적인 차이를 보이지 않았다. Glycine은 소장 단백질과 DNA에 있어서 대조군보다는 높고 Nuc군보다는 낮은 경향을 보여 주었으나 [$^3$H]-thymidine incorporation은 누공과 비누공부분에서 모두 가장 높게 나타났으며 특히 비누공부분에서는 유의적으로 증가되어 있었다. Glutamine의 효과는 glycine 투여 효과와 비슷하거나 오히려 떨어지는 것으로 나타났다.

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in vitro에서 B(a)P이 착상전 마우스 배자에 미치는 독성학적 영향에 관한 연구 (Toxicological Effects of B(a)P on Preimplantation Mouse Embryos in Vitro)

  • 박귀례;이유미;김판기;신재호;강태석;김주일;장성재
    • 한국환경보건학회지
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    • 제24권2호
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    • pp.126-133
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    • 1998
  • Effects of B(a)P on preimplantation mouse embryos in vitro were studied. Preimplantation mouse embryos were exposed to a concentration of 0.3, 1, 3 and 10 $\mu$M B(a)P for 72 hrs. The toxicological effects of B(a)P were evaluated by morphological observation of embryos up to the blastocyst stage, and by measuring DNA, RNA and protein synthesis by radioactive precursor incorporation. At 1 $\mu$M B(a)P did not affect preimplantation development but interfered with hatching and ICM formation. Suppressing effect of ICM formation was dose dependent. At the eight cell stage, the developmental rate was decreased at above 3 $\mu$M of B(a)P. At the blastocyst stage, attachment and trophoblast outgrowth were diminished at the 10 $\mu$M of B(a)P and ICM formation was decreased at 1 $\mu$M of B(a)P. Inner cell number of blastocyst was decreased dose dependently. So, number of ICM was one of the most sensitive and toxicological end point. The RNA incorporation rate of 0.1 $\mu ^3$H-uridine was dosedependent and the protein incroporation of 0.5 $\mu Ci ^{35}$S-methionine showed a significant decrease after 48 hrs. But the DNA incorporation rate of methyl-$^3$H thymidine was not affected. Our results suggested that B(a)P did not affect the DNA replication but transcription was inhibited by dose dependent manner. There delay of development during the blastocyst stage was mainly due to the inhibition of RNA synthesis followed by protein synthesis.

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호수 생태계에서 세균 생산량 측정 방법의 비교 (Comparison of Methods for the Measurement of Bacterial Production in a Lake Ecosystem)

  • 김명운;강찬수;김상종
    • 미생물학회지
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    • 제28권4호
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    • pp.318-323
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    • 1990
  • The bacterial secondary production was measured at 6 sites of Lake Soyang in October, 1989 by $^{3}$H-thymidine incorporation rate. Verfication for the method of bacterial secondary production measurement showed that $^{3}$H-thymidine incorporated into DNA, RNA and protein by average percentage of 38.45, 42.27 and 20.07%, respectively. THe more increased incoporated $^{3}$H-thymidine, the more increasde DNA fraction, but protein fraction was generally low. Incorporation of rate of /usp 3/H-thymidine. $^{3}$H-leucine into protein correlated with protein fraction of incorporated $^{3}$H-thymidine. Conversion factors were calculated as follows; $1.83*10 ^{20}$ cells/moles of thymidine incorporated/hr and 1.69*10$^{22}$ cells/moles of leucine incorporated/hr.

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Inhibitory Effect of Scutellaria barbata Don Water-extracts on Growth and DNA Incorporation of Human Cancer Cells

  • Kim, Dong-Il
    • 대한한의학회지
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    • 제27권4호
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    • pp.162-173
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    • 2006
  • The water-extracts of Scutellaria barbata Don (SBDE) were isolated from Chinese medicinal plant sources. The extracts showed strong growth-inhibitory activity and cancer chemopreventive activity on the growth and DNA incorporation of MG63 human osteosarcoma and K562 human leukemia cell lines. The growth of human cancer cells was inhibited in the presence of the extracts (20, 50 and 100 ${\mu}$g/ml), and the effects were concentration-dependent and incubation time-dependent up to 8 days. When 50 ${\mu}$g/ml of the extracts was added to the media of MG63 and K562, cell growth after 8 days or 6 days of incubation was retarded by 93.2 to 97.3% of the control group. Morphological changes of MG63 and K562 cell lines were observed. As the concentration of the extracts increased up to 50 ${\mu}$g/ml, degree of cell aggregation decreased. Moreover, the DNA incorporation of the cells which were labeled with [3H] thymidine was significantly reduced after 3 days of incubation at $37^{\circ}C$ with the extract. Therefore, it is suggested that the extract is highly effective on inhibition of cancer cell growth. The extract also inhibited gene expression of IGF-II in transcriptional level. Since IGF-II works as a mitogenic effector on MG63 and K562 cell lines, these results suggest that the growth inhibition is in part mediated through the inhibition of IGF-II gene expression.

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쥐에서 임신기, 수유기 및 이유후에 식이로 섭취한 어유가 뇌조직의 DHA 분포량에 미치는 영향 (Influence of Dietary Supplementation of Fish Oil at Different Life Cycle on the Incorporation of DHA into Brain in Rats)

  • 박기호;박현서
    • Journal of Nutrition and Health
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    • 제31권7호
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    • pp.1100-1111
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    • 1998
  • The incorporation of docosahexaenoic acid(DHA) and arachidonic acid(AA) into brain and liver lipid has been compared in male pups from binth to 10 weeks old by feeding DHA-rich experimental diets or chow diets to dams from pregnancy in rats. The experimental DHA-rich diets contained 7g fish oil and 3g corn oil per 100g diet. There were three experimental groups, FO-I : Dams were fed DHA-rich diet during pregnancy and lactation, and their it pups fed the same diet until 10 weeks old. FO-II Dams fed chow diet during pregnancy and DHA-diet during lactation, and their pups fed the same DHA-diet until 10 weeks. FO-III : Dams fed chow diet during gestation and lactation, and then the pups fed DHA-diet after weaning. The relative % of DHA in hepatic lipid was about 12% with chow diets, but increased rapidly to 20-25% level when DHA-rich diets were supplied after weaning. The AA(%) of FO-III group was relatively high when a chow diet containing higher amount of linoleic acid was given, but there was no significant difference between the groups after feeding on a DHA-rich diet. When the DHA-rich diet was supplied from pregnancy(FO-I), the relative % of DHA in brain lipid was 13.7% at birth and continuously increased to a maximum level(17.2%) at 3-weeks and then was sustained until 5 weeks old. Similar levels of DHA incorporation were observed when DHA-rich diet was supplied from lactation(FO-II). However, the pups of FO-III group showed significantly lower levels of DHA incorporation(72%) at birth. These livels slowly increased and reached an 87% level of FO-I at 10 weeks when the pups ate DHA-rich diets after weaning. The relative % of AA in brain lipid was 10.4% in the FO-I group at birth, which was significantly lower than those of other groups, but there was no significant difference between groups after feeding DHA-rich diets in all groups. The Ah(%) level increased to maximum(11-12%) at 3-weeks and then was slightly reduced and was sustained at about 10% after S-weeks. Total amounts of DNA in the whole brain rapidly reached maximum level at 3-weeks and then was sustained at a constant level after S-weeks. DNA content was not significantly different between groups at birth, but it was significantly higher in FO-I and FO-II groups than in FO-III group at 3-weeks. However, DNA content in FO-III group was continuously increased to 80% level of FO-I at 10-weeks after feeding DHA-rich diet since weaning. In conclusion, the DHA(%) in whole brain was most effectively deposited when DHA-rich diet had been supplied during pregnancy and lactation in rats. However, DHA supplementation after weaning also improved the incorporaton of DHA into brain and content of DNA even though brain development was almost completed, which suggests that DHA supplementation might be necessary to improve brain development in humans during infancy as well as pregnancy and lactation. (Korean J Nutrition 31(7) 1100-1111, 1998)

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