• Title/Summary/Keyword: DNA-cleavage

검색결과 388건 처리시간 0.022초

HL-60 세포에서 Camptothecin의 apoptosis 유도작용 (Induction of Apoptosis by Camptothecin in HL-60 Cells)

  • 김해종;천영진;김미영
    • 약학회지
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    • 제43권3호
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    • pp.385-390
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    • 1999
  • Camptothecin (CPT) has been known to induce apoptosis in various cancer cell lines. To examine the intracellular apoptotic death signal initiated by CPT, we investigated the possible connection between caspase-3 activation and GSH depletion during CPT-induced apoptosis in HL-60 cells. Treatment of cells with $1{\;}{\mu}M$ CPT induced PARP cleavage accompanied by DNA fragmentation. z-VAD-fmk, a caspase-3 inhibitor, blocked the CPT-induced DNA fragmentation. Pretreatment of cells with N-acetylcysteine, a precursor of GSH biosynthesis, failed to inhibit CPT-induced PARP celavage and DNA gragmenatation. No significant changes in GSH depletion is not essential for caspase activation during CPT-induced apoptosis. We also investigated whether CPT-induced apoptosis is associated with changes of the levels of Bax and Bcl-2, two proteins involved in the control of apoptosis. Bcl-2 levels exhibited a late decrease compared with the kinetics of DNA fragmentation, whereas Bax levels increased more rapidly after CPT treatment. These results suggest that Bax plays more important role than Bcl-2 in inducing DNA fragmentation and may function upsteam of proteolytic activation of caspase-3 pathway in CPT-induced apoptosis.

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박테리오파아지 P2의 sir 변이를 억제하는 새로운 박테리오파아지 P4 유도체인 P4 ost2의 분리와 동정 (Isolation and Identification of the Bacteriophage P4 ost2, Suppressing sir Mutations of Bacteriophage P2)

  • 김경진
    • 미생물학회지
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    • 제39권4호
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    • pp.277-282
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    • 2003
  • P2의 sir 변이를 억제하는 변이체 박테리오파아지 P4를 얻기 위하여, P4 ash8 sid71 kmr intS를 박테리오파아지 P2 sir3 용원소에 plating하여 플라크를 형성하는 P4 ost2를 분리하였다. P4 ost2는 1단계 생장실험에서 P2의 sir 변이를 억제하는 P4 변이체로 나타났다. 제한효소 절단 반응으로 유전체 DNA의 재배열이 일어난 단편을 찾아 cloning과 sequencing을 거쳐 P4 ost2의 구조를 규명하였다. P4 ost2의 DNA는, 6.9 kb의 결실을 가진 P4 ash8 sid71 kmr intS의 불완전한 trimer로 2개의 cos-cleavage 부위를 가지는 28.8 kb의 유전체로 밝혀졌다. CsCl 부양균등밀도 편차실험으로 P4 ost2의 DNA 크기를 확인하였고 파아지 머리 안에 packaging된 DNA에 대한 기초적인 구조를 파악하였다. P4 ost2의 분리 동정을 통해, P4 유전체 상의 cos-cleavage 부위의 개수가 Sir-type 파아지 머리의 packaging 반응과는 관련이 없다고 추정할 수 있었다.

미선나무 잎 추출물의 항산화 및 산화적 DNA 손상억제 활성 (Antioxidant Activities and Inhibitory Effect on Oxidative DNA Damage of extracts from Abeliophylli distichi Folium)

  • 박재호
    • 대한본초학회지
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    • 제26권4호
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    • pp.95-99
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    • 2011
  • Objective : In this study, we demonstrate the protective effect on oxidative DNA damage of leaf extracts from Abeliophylli distichi Folium via its antioxidant activity for the establishment of new value for the herbal medicine. Methods : Abeliophylli distichi Folium leaves were extracted with hot-water and ethylacetate (EtOAC). The 1,1-diphenyl-2- picrylhydrazyl (DPPH) radical and hydroxyl scavenging assay and $Fe^{2+}$ chelating assay were performed for antioxidative effect and ${\varphi}$X-174 RF I DNA cleavage assay and intracellular DNA damage assay were used for inhibitory effect of intracellular DNA damage. Results : In DPPH, Hydroxyl radical scavenging activity and $Fe^{2+}$ chelating activity of EtOAC extracts were 94.72%, 62.88%, 41.13%, and hot-water extracts were 88.86%, 56.7%, 37.4% at 200 ${\mu}g/m{\ell}$, respectively. Also, those extracts showed protective effect of DNA damage against the oxidative stress. Conclusion : These results indicated that the leaf extracts of Abeliophylli distichi Folium can be used as a natural antioxidants, which effectively inhibits the oxidative DNA damage.

Chlamydomonas reinhardtii에서 순화한 Endonuclease의 특징과 Polyamine의 영향 (Characterization of and Polyamine Effect on Endonuclease from Zygotes of Chlamydomonas reinhardtii)

  • 김재윤
    • Journal of Plant Biology
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    • 제33권4호
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    • pp.293-301
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    • 1990
  • We have purified and characterized a deoxyribonuclease from zygotes of the eukaryotic green alga, Chlamydomonas reinhardtii and investigated effects of the polyamines, putrescine, spermidine and spermine on the purifed endonuclease-catalyzed cleavage of plasmid DNA. The enzyme has a molecular weight of about 37 kDa as measured by gel filtration and SDS-polyacrylamide gel electrophoresis. There is no requirement for a divalent cation. The activity is sensitive to ionic strength, as NaCl and KCl result in inhibition. The cleavage of plasmid DNA by the purified endonuclease was effectively inhibited by polyamines. The enzyme activity was inhibited more effectively by spermine than by spermidine. The inhibition by putrescine was lower than the other two polyamines.

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Drosophila robusta species group 2종 (D.lacertosa 와 D.sordidula)의 mtDNA 변이에 의한 종분화정도

  • 최준길;박제철
    • Animal Systematics, Evolution and Diversity
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    • 제11권4호
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    • pp.469-477
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    • 1995
  • Drosophila virilis section 중 D. robusta 종군내에서 D. lacertosa 아군의 D. lacertosa 와 D. robusta 아군의 D.sordidula 에 대한 형태학적 특징 및 세포학적 특징(핵형분석)간에 차이가 크기 때문에 이 두종을 대상으로 종의 분화정도와 종형성 과정을 알아보기 위한 연구의 일환으로 10가지 제한효소를 사용하여 mtDNA 의 절단인식부위를 분석하였다. 그 결과, D.lacertosa 와 D.sordidula 의 전체 genome size는 공히 15.7kbp 인 것으로 나타났으며, mtDNa 의 제한효소 fragment 수는 각각 26개와 32개인 것으로 조사되었다. 또한 2 종류의 제한효소를 동시에 처리하여 제한효소 지도를 작성하여 보았을 때, 이들 두종의 제한효소 지도의 형태에서 매우 큰 차이가 있는 것으로 나타났다. 같은 종군내에서 형태학적 , 세포학적 차이 및 mtDNA 의 제한효소 지도에 의한 차이로 볼 때 이들 2종의 차이는 아군수준으로까지 분류할 수 있을정도로 두종의 분화가 오래전에 이루어졌음을 시사하였다.

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Roles of the meta- and the ortho-Cleavage Pathways for the Efficient Utilization of Aromatic Hydrocarbons by Sphingomonas yanoikuyae Bl

  • 송정민;김영민;Gerben J. Zylstra;김응빈
    • 미생물학회지
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    • 제38권4호
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    • pp.245-245
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    • 2002
  • Catabolic pathways for the degradation of various aromatics by Sphingomonas yanoikuyae Bl are intertwined, joining at the level of substituted benzoates, which are further degraded vita ring cleavage reactions. The mutant strain EK497, which was constructed by deleting a large DNA region containing most of the genes for biphenyl, naphthalene, m-xylene, and m-toluate degradation, was unable to grow on all of the aromatics tested except for benzoate as the sole source of carbon and energy.S. yanoikuyae EK497 was found to possess only catechol ortho-ring cleavage activity due to deletion of the genes for the meta-cleavage pathway. Wild-type S. yanoikuyae Bl grown on benzoate has both catechol orthoand meta-cleavage activity. However, m-xylene and m-toluate, which are metabolized through methylbenzoate, and biphenyl, which is metabolized through benzoate, induce only the meta-cleavage pathway, suggesting the presence of a substrate-dependent induction mechanism.

Human Topoisomerase I-DNA 절개가능 복합체에 대한 Indenoisoquinoline 유도체들의 결합양상 연구 (Binding Mode Studies of Indenoisoquinoline Analogues into Human Topoisomerase I-DNA Complex Using Flexible Docking)

  • 박인선;김보연;김춘미;최선
    • 약학회지
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    • 제53권4호
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    • pp.228-234
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    • 2009
  • Topoisomerase I (Topo I) participates in the DNA replication, transcription, and repair. Binding of Topo I inhibitor to the Topo I-DNA cleavage complex forms stabilized ternary complex which blocks DNA religation and ultimately causes cell death. Camptothecin (CPT) and its derivatives have been among the most effective anticancer drugs by inhibition of topo I. However, efforts to synthesize non-CPT drugs have been actively going on because the CPT derivatives have several limitations such as poor solubility, short half-life, and side effects. As an indenoisoquinoline, NSC314622 is not as potent as CPT, but its chemical stability and slower reversibility of the cleavage complex made it a good lead compound. Recently, a series of indenoisoquinoline analogues were synthesized with substituted dimethoxy or methylenedioxy on the aromatic ring and alkylamino on the lactam nitrogen. Some of them showed quite good Topo I inhibitory activity. Using the computer docking program, Surflex-Dock, indenoisoquinoline analogues were docked into the human Topo I-DNA cleavable complex. The docking results showed that the compounds with activity better than NSC314622 intercalated between the -1 and +1 base pairs at the cleavage site, but those with little or no activities did not appear to intercalate. These results could be useful to design new Topo I inhibitors improved than CPT.

Pseudomonas sp. strain DJ77로 부터 phenanthrene 분해 유전자군의 클로닝과 대장균에서의 발현 (Molecular Cloning of a Gene Cluster for Phenanthrene Degradation from Pseudomonas sp. Strain DJ77 and Its Expression in Escherichia coli)

  • 김영창;윤길상;신명수;김흥식;박미선;박희진
    • 미생물학회지
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    • 제30권1호
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    • pp.1-7
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    • 1992
  • Pseudomonas sp. DJ77 의 chromosomal DNA로 부터 6-8kb XhoI 절편 상에 존재하는 phenanthrene 분해에 관련된 유전자군을 vector pBLUESCRIPT SK(+)에 클로닝하였다. 이렇게 얻은 재조합 plasmid인 pHENX7을 가지고 있는 JM101 균주는 3-methylcatechol을 노란색의 meta-cleavage 화합물로 전환학 수 있었다. 그러자 삽입된 절편의 방향이 반대가 되도록 제조한 pHENX7R 은 extradiol dioxygenase 활성을 나타내지 않기 때문에 전사방향을 알 수 있었다. pHENX7과 이의 유도체들을 지니는 JM101 균주에서 PhnC(24kDa), PhnD(31 kDa), PhnE(34 kDA), PhnF(15 kDa) 의 4 polypeptide 를 확인학 수 있었고 개개의 유전자의 위치와 범위를 알 수 있었다. 유전자 순서는 phnC-phnD-phnE-phnF-phnG 이었으며, phnD, phnE, phnG 는 각기 gluthione S-transferase, meta-cleavage compound hydrolase, extradiol dioxygenase, metacleavage compound dehydrogenase 의 유전자이었다.

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