• Title/Summary/Keyword: DNA condensation

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Condensation of DNA by a Histone-like Protein in Escherichia coli

  • Kim, So-Youn;Hwang, Deog-Su
    • BMB Reports
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    • v.28 no.2
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    • pp.143-148
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    • 1995
  • In E. coli, chromosomal DNA associated with proteins is condensed into an organized structure known as nucleoid. Using a nitrocellulose filter binding assay to identify proteins forming nucleoid, a 21 kDa protein was purified from E. coli. The molecular weight of the purified protein was 21 kDa on SDS-polyactylamide gel electrophoresis and 24 kDa on gel permeation chromatography. A molecular weight of 21 kDa on SDS-polyacrylamide gel electrophoresis is unique among known proteins which are believed to be involved in the formation of nucleoid in E. coli. The 21 kDa protein nonspecifically binds to both double-stranded and single-stranded DNA. Sedimentation in a sucrose gradient revealed that the protein induced significant condensation of both supercoiled plasmid DNA and linear bacteriophage $\lambda$ DNA On the basis of quantitative Western-blot analysis, approximately 40,000 molecules of the protein were estimated to exist in an E. coli. The biochemical properties and cellular abundance of the 21 kDa protein suggest that this protein participates in the formation of nucleoid in E. coli.

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Ordered Fragmentation of pDNA induced by PEG-PLL block copolymer -Correlation between Condensation degree and Biological Activity by Cell-Free System-

  • Osada, Kensuke;Doi, Motoyoshi;Shiotani, Tomonori;Yamasaki, Yuichi;Kataoka, Kazunori
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.254-254
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    • 2006
  • The sensitivity of plasmid DNA (pDNA) to S1 nuclease, an enzyme to cleave a single-strand DNA, was dramatically modulated through a supramolecular assembly (polyion complex micelle) with a synthetic block copolymer, poly(ethylene glycol)-b-poly(L-lysine) (PEG-PLL). The pDNA condensed in stoichiometric charge ratio was cleaved into 7 fragments each being 10/12, 9/12, 8/12, 6/12, 4/12, 3/12, and 2/12 of the original DNA length, on the other hand, the pDNA condensed in higher charge ratios (>4), were digested into non-specific manner. Condensation of the pDNA was investigated from two viewpoints that how does the rigid DNA molecules fold and condense and how does the condensation influence their biological activity.

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Polyplex Formation of Calf Thymus DNA with Branched and Linear Polyethyleneimine

  • Han, Jung-Ho;Kim, Seog K.;Cho, Tae-Sub;Lee, Jae-Cheol;Joung, Hyun-Sook
    • Macromolecular Research
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    • v.12 no.5
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    • pp.501-506
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    • 2004
  • We have investigated the spectral properties of DNA, including its electric absorption, circular and linear dichroism (CD and LD), and fluorescence emission, in the DNA-linear polyethyleneimine (LPEI) and DNA-branched polyethyleneimine (BPEI) complexes at various polymer concentrations. The spectral properties of both complexes are similar. We observed a relatively moderate change in the absorption and CD spectra at low amine/DNA phosphate (NIP) ratios (< 0.5), followed by a drastic collapse within the N/P range from 0.8 and 1.0. The absorption and CD spectra recovered as the N/P ratio increased to ca. 1.2. In contrast, the LD and emission of ethidium intercalated between the DNA bases decreased almost linearly at N/P ratios between 0.0 and 1.0. These spectra never recovered at higher N/P ratios. We believe that the moderate changes in the spectrum at low N/P ratios occurred because of electrostatic interactions between DNA and BPEI, while the collapsed spectra at N/P ratios between 0.5 and 1.5 occurred because of condensation/aggregation of the DNA. Considering the structure of the polymers, we suggest that the secondary amino group of LPEI and all three amino groups of BPEI are equally involved in DNA condensation.

Cytopathic Effects of Japanese Encephalitis Virus Structural Proteins in BHK-21 Cells (BHK-21 세포에서의 일본뇌염바이러스 구조단백질에 의한 세포독성)

  • 성기민;정용석
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.213-220
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    • 2002
  • Inducible expression system for the three structural proteins, capsid (C), precursor membrane (prM/M), and envelop (E) of Japanese encephalitis virus (JEV) was established in BHK-21 cells. Doxycycline, a tetracycline analog, was utilized as an inducer. Transfectants BHK-21/IV (vector only), BHK-21/IC (for C), BHK-21/IP3 (for prM), and BHK-21/IE1 (for E) were selected and cloned in the presence of G4l8 or hygromycin. Transcribed mRNAs for the corresponding genes were observed after doxycycline induction. Effects by the JEV structural gene expression on the transfectants were monitored via cell growth, chromatin condensation, internucleosomal DNA fragmentation, and DNA contents analyses. Clear cell growth retardation and chromatin condensation were observed in all three transfectants while only BHK-2/IC corresponded to the induction status in the DNA fragmentation and DNA content analyses. Combined results, therefore, suggested that JEV capsid protein should be one of the direct and independent factors in apoptotic cell death induced by IEV infection.

The utility of sperm DNA damage assay using toluidine blue and aniline blue staining in routine semen analysis

  • Kim, Hee-Sun;Kang, Moon Joo;Kim, Sung Ah;Oh, Sun Kyung;Kim, Hoon;Ku, Seung-Yup;Kim, Seok Hyun;Moon, Shin Yong;Choi, Young Min
    • Clinical and Experimental Reproductive Medicine
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    • v.40 no.1
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    • pp.23-28
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    • 2013
  • Objective: The aim of the present study was to examine the relationship among male age, strict morphology, and sperm chromatin structure and condensation. Methods: Sperm samples from a total of 100 men underwent semen analysis, and sperm chromatin structure and condensation were assessed with toluidine blue (TB) and aniline blue (AB) tests. Results: Prevalence of strict morphology of less than 4%, and abnormal sperm chromatin structure and condensation did not show any statistically significant differences according to male age (p=0.605, p=0.235, and p=0.080). No significant correlation was demonstrated among age of male partners, strict morphology, and abnormal sperm chromatin structure using TB and AB tests. However, abnormal sperm chromatin condensation was positively associated with sperm chromatin structure (r=0.594, p=0.000) and showed negative correlation with strict morphology (r=-0.219, p=0.029). Conclusion: The tests for sperm chromatin condensation showed a significant association with strict morphology. Further study is needed to elucidate the relationship between clinical outcome and sperm chromatin tests.

Up-regulation of Bax is associated with DNA topoisomerase I inhibitor β-lapachone-induced apoptosis in human prostate carcinoma cells (DNA topoisomerase I 억제제 β-lapachone에 의한 전립선 암세포의 성장억제 기전연구)

  • 공규리;최병태;최영현
    • Journal of Life Science
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    • v.12 no.4
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    • pp.469-476
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    • 2002
  • The DNA topoismerase I inhibitor $\beta$-lapachone, the product of a tree from South America, is known to exhibit various biological properties, however the mechanisms of which are poorly understood. In the present report, we investigated the effects of $\beta$-lapachone on the growth of human prostate carcinoma DU-145 cells. Upon treatment with $\beta$-lapachone, a concentration-dependent inhibition of cell viability was observed and cells developed many of the hallmark features of apoptosis, including condensation of chromatin and DNA fragmentation. Flow cytometry analysis confirmed that $\beta$-lapachone increased populations of apoptotic-sub Gl phase. In addition, proteolytic cleavages of poly (ADP-ribose) polymerase (PARP) and $\beta$-catenin protein were observed after treatment of $\beta$-lapachone. These apoptotic effects of $\beta$-lapachone in DU-145 cells were associated with marked induction of Bax protein, however the levels of Bcl-2 expression were decreased in a dose-dependent manner.

Effect of Proliferation Inhibition on B16/F10 Melanoma Cell by Chloroform Extract from Cornis fructus (산수유 클로로포름 추출물에 의한 B16/F10 melanoma세포의 증식억제효과)

  • 최원형;천현자;백승화;우원홍
    • Biomolecules & Therapeutics
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    • v.11 no.2
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    • pp.151-156
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    • 2003
  • Cornis fructus have various biological effects and major chemical components have been tannins, saponins, ursolic acids, gallic acids, linoleic acids, morronisides, cornins and loganins. The main aim of the present study is measurment the effect of chloroform extract from Cornis fructus on proliferation inhibition and Cell death. Cells were cultured in the presence of chloroform extracts from Cornis fructus for 48 h. after 48h treatment of B16/F10 melanoma cells with chloroform extracts, the cells were observed a dose-dependent inhibitions of cell viability with cell death in their proliferation. the cells were estimated cell viability, cell number, total DNA fragmentation and chromatin condensation in a dose-dependent manner. It also caused cell death as measured by cell morphology, DNA fragmentation and nucleus chromatin condensation. therefore, these results suggest that chloroform extracts from C. fructus is inhibitory proliferation and is related to cell death in this cells.

Protective Effects of Chitosan on the Cadmium Cytotoxicity in Rat Glioma Cells (흰쥐 신경교종세포에서 카드뮴 세포독성에 대한 키토산의 효과)

  • 백용아;이정래;김강득;김혜원;이한솔;허정무;오재민;최민규;정연태
    • 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.

Condensation-Decondensation Structural Transition of DNA Induced by Reversible Ligand Binding : Effect of Urea on Anomalous Absorbance-Temperature Profile of Spermine-DNA Complex (可逆的 리간드 結合에 의하여 誘發되는 DNA의 응축-풀림 構造變移 : Spermine-DNA 複合體의 異例的 吸光度-溫度 樣相에 미치는 Urea의 影響)

  • Thong-Sung Ko;Chan Yong Lee
    • Journal of the Korean Chemical Society
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    • v.29 no.5
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    • pp.533-538
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    • 1985
  • To investigate the importance of the hydrophobic interaction in the spermine-induced collapse of DNA to a compact structure, the effect of urea on the anomalous absorbance-temperature profile of calf thymus DNA has been investigated. With the increase of the urea concentration, the trough phase of the anomalous absorbance-temperature profile was eliminated eventually. The cooperativity, enthalpy, and the midpoint of the transition to the trough region are more sensitive to urea than those of the Tm-region transition. The present data of the adverse effect of urea, a hydrophobic environmental reagent, on the thermal stabilization of the condensed state of DNA, suggest that hydrophobic interaction may play an important role in the stabilization of the tertiary structure of the collapsed state of DNA.

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Hypersensitive and Apoptotic Responses of Pepper Fruit Against Xnthomonas axonopodis pv. glycines Infection

  • Chang, Sung-Pae;Kim, Young-Ho
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.72.1-72
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    • 2003
  • Generally, plants defend themselves against pathogens by structural and biochemical reactions. Defense structures act as physical barriers and inhibit the pathogen from gaining entrance and spreading through the plant. Xanthomonas axonopodis pv glycines, the causal pathogen of bacterial pustule of soybean, causes hypersensitive response (HR). When pepper fruits were inoculated with X. axonopodis pv. glycines, in situ, time-series defense-related structural changes occurred in the inoculated sites. Early responses were programmed cell death (PCD), characterized by condensation and vacuolization of the cytoplasm, condensation of nuclear materials, and fragmentation of the nuclear DNA, which were observed by transmission electron microscopy. Nuclear fragmentation was proven by TUNEL method under confocal laser scanning microscopy and DNA laddering through eletrophoresis. At later stages, plant responses were cell elongation and cell division, forming a periderm-like boundary layer that demarcated healthy tissues from the inoculation sites. Using several stains such as toluidine blue, sudan IV, annexin V, and phloroglucinol-HCl, defense-related materials and structural changes were also examined.

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