• Title/Summary/Keyword: cleavage.

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Species-Specific Cleavage by RNase E-Like Enzymes in 5S rRNA Maturation

  • RYOU SANG-MI;KIM JONG-MYUNG;YEOM JI-HYUN;KIM HYUN-LI;GO HA-YOUNG;SHIN EUN-KYOUNG;LEE KANGSEOK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1100-1105
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    • 2005
  • Previous work has identified a Streptomyces coelicolor gene, rns, encoding a 140 kDa protein (RNase ES) that exhibits the endoribonucleolytic cleavage specificity characteristic of RNase E and confers viability on and allows the propagation of E. coli cells lacking RNase E. Here, we identify a putative S. coelicolor 9S rRNA sequence and sites cleaved by RNase ES. The cleavage of the S. coelicolor 9S rRNA transcript by RNase ES resulted in a 5S rRNA precursor (p5S) that had four and two additional nucleotides at the 5' end and 3' ends of the mature 5S rRNA, respectively. However, despite the similarities between RNase E and RNase ES, these enzymes could accurately process 9S rRNA from just their own bacteria, indicating that these ancient enzymes and the rRNA segments that they attack appear to have co-evolved.

The Nucleophilic Crown Ether Ring Cleavage of Nitro Derivatives of Dibenzo-18-crown-6-(I) (친핵성 치환 반응에 의한 Dibenzo-18-crown-6 의 Nitro 유도체 화합물의 Crown Ether 고리 끊어짐 (제 1 보))

  • Sae Hee Chang;Sung Ah Cho
    • Journal of the Korean Chemical Society
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    • v.32 no.1
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    • pp.71-78
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    • 1988
  • The crown ether ring of 20, 21, 24, 25-tetranitrodibenzo-18-crown-6(TNDB-18-C-6) was cleaved by various alcoholic bases to give 2,4,5-trialkoxynitrobenzene derivatives and 4,5-dialkoxy-1,2-dinitrobenzene derivatives as the major products, and bis[(alkoxynitrophenoxy)ethyl]ether derivatives from partially cleaved crown ether ring as the minor products. The ring cleavage reaction of TNDB-18-C-6 with ethylene glycolic base resulted ring contraction through intramolecular nucleophilic cyclization of the initially formed ring cleavage product to give nitro derivatives of DB-14-C-4.

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Density Functional Study on the C-H Bond Cleavage of Aldimine by a Rhodium(I) Catalyst

  • Yoo, Kyung-Hwa;Jun, Chul-Ho;Choi, Cheol-Ho;Sim, Eun-Ji
    • Bulletin of the Korean Chemical Society
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    • v.29 no.10
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    • pp.1920-1926
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    • 2008
  • We investigated the C-H bond activation mechanism of aldimine by the [RhCl$(PPH_3)_3$] model catalyst using DFT B3LYP//SBKJC/6-31G*/6-31G on GAMESS. Due to their potential utility in organic synthesis, C-H bond activation is one of the most active research fields in organic and organometallic chemistry. C-H bond activation by a transition metal catalyst can be classified into two types of mechanisms: direct C-H bond cleavage by the metal catalyst or a multi-step mechanism via a tetrahedral transition state. There are three structural isomers of [RhCl$(PH_3)_2$] coordinated aldimine that differ in the position of chloride with respect to the molecular plane. By comparing activation energies of the overall reaction pathways that the three isomeric structures follow in each mechanism, we found that the C-H bond activation of aldimine by the [RhCl$(PH_3)_3$] catalyst occurs through the tetrahedral intermediate.

Expression of Poly (ADP-ribose) Polymerase During Apoptosis Induced by Ultraviolet Radiation in HeLa $S_3$ Cells

  • Chang, Jeong-Hyun;Kwon, Heun-Young
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.349-354
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    • 2006
  • Induction of apoptosis allows the organism to get rid of abnormal cells and also of tumor cells. Understanding the mechanism involved in Ultraviolet radiation (UV) induced apoptosis may improve its therapeutic efficacy. In this study, we present expression of poly (ADP-ribose) polymerase (PARP) during apoptosis induced by UV in HeLa $S_3$ cells. Four different assays were performed in this study: morphological assessment of apoptotic cells and cell viability, DNA fragmentation analysis by agarose gel electrophoresis, quantitative assay of fragmented DNA, and expression of PARP by the western blot analysis. The percentages of apoptotic HeLa $S_3$ cells irradiated with $75J/m^2$ UV was increased continuously from 3 hrs incubation. DNA ladder pattern was appeared at 6 hrs. The amount of nucleosomal DNA fragments in cells treated UV increased from 3 to 12 hrs incubation and gradually decreased. The cleavage of PARP in HeLa $S_3$ cells irradiated with UV was induced, and the cleavage of PARP was more delayed in the cells pretreated with $5J/m^2$ UV and subsequently irradiated with $75J/m^2$ UV. than that in the cells only irradiated with $75J/m^2$ UV. Thus these data suggest that the cleavage of PARP relates with DNA fragmentation associated with apoptosis.

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Recombination Activating Gene 1 Product Alone Possesses Endonucleolytic Activity

  • Kim, Deok-Ryong
    • BMB Reports
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    • v.36 no.2
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    • pp.201-206
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    • 2003
  • Two lymphoid-specific proteins, RAG1 and RAG2, are required for the initiation of the V(D)J recombination in vitro. The V(D)J cleavage that is mediated by RAG proteins at the border between the coding and signal sequences results in the production of a hairpin at the coding end and a double-stranded break at the signal end. Two hairpin coding ends are re-opened, modified, and sealed; whereas, the signal ends are directly ligated. Here I report that only RAG1 can carry out a distinct endonucleolytic activity in vitro using an oligonucleotide substrate that is tethered by a short single-stranded DNA. The purified RAG1 protein alone formed a nick at the near position to the recombination signal sequence. This endonucleolytic activity was eliminated by immunoprecipitation using the RAG1-specific antibody, and required the 3'-hydroxy group. All of the RAG1 mutants that were incapable of the nick and hairpin formation in the V(D)J cleavage analysis also showed this new endonucleolytic activity. This suggests that the nicking activity that was observed might be functionally different from the nick formation in the V(D)J cleavage.

Antioxidant Compounds of Oryza sativa L (벼의 항산화성분)

  • Min, Byung-Sun;Lee, Hyeong-Kyu;Zee, Ok-Pyo;Moon, Hyung-In
    • Korean Journal of Pharmacognosy
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    • v.33 no.3 s.130
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    • pp.173-176
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    • 2002
  • In search for the plant-derived antioxidant compound, it was found that the EtOAc, BuOH extracts obtained from the leaves parts of Oryza sativa L. which exhibited a significant antioxidant activity from 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical cleavage. Activity-guided fractionation on the basis of the inhibitory activity upon the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical cleavage. and repeated column chromatography afforded several antioxidant compounds from Oryza sativa L. The structures and stereochemistry of these compounds were established on the basis of analysis of spectra and some chemical transformations as follows: 5,7-dihydroxy- 8-methoxyflavone, $acacetin-7-O-{\beta}-rutinoside$, $pectolinarigenin-7-O-{\beta}-rutinoside$. At antioxidant activity test for isolated three compounds, antioxidant activity was showed too.

Effect of Gonadotropin Administration on the Timing of Ovulation, Fertilizable Life of Eggs and Cleavage of Embryos in Rabbit (성선자극 호르몬 투여가 토끼의 배란시간, 난자의 수정능력 보유시간과 난할 속도에 미치는 영향)

  • 김병기
    • Journal of Life Science
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    • v.8 no.5
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    • pp.576-581
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    • 1998
  • The present study was carried out to investigate the effect of gonadotropin administration on the timing of ovula-tion, fertlizable life of eggs and cleavage of embryos in rabbit. Mature angora rabbits were primed for superovulation with PMSG 100IU. Eighty hours later, the rabbit were induced to ovulate with HCG 100IU. Ovulation had started at 10hours after HCG injection and finished at about 16hours. Fertilizable life of eggs were lasted for 8hours after ovulation. The most frequent developmental stage observed from the embryos recovered at 24, 48, 72, 96, and 120 hours after HCG injection was 2-ceIL, 16-cell, morula, blastocyst and blastocyst, respectively.

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Development of Cleavage Fracture Toughness Locus Considering Constraint Effects

  • Chang, Yoon-Suk;Kim, Young-Jin;Ludwig Stumpfrock
    • Journal of Mechanical Science and Technology
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    • v.18 no.12
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    • pp.2158-2173
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    • 2004
  • In this paper, the higher order terms in the crack tip stress fields are investigated macroscopically for more realistic assessment of structural material behaviors. For reactor pressure vessel material of A533B ferritic steel, effects of crack size and temperature have been evaluated using 3-point SENB specimens through a series of finite element analyses, tensile tests and fracture toughness tests. The T-stress, Q-parameter and q-parameter as well as the K and J-integral are calculated and mutual relationships are investigated also. Based on the evaluation, it has proven that the effect of crack size from standard length (a/W=0.53) to shallow length (a/W=0.11) is remarkable whilst the effect of temperature from -20$^{\circ}C$ to -60$^{\circ}C$ is negligible. Finally, the cleavage fracture toughness loci as a function of the promising Q-parameter or q-parameter are developed using specific test results as well as finite element analysis results, which can be applicable for structural integrity evaluation considering constraint effects.

Autoxidation Products of Phytofluene in Liposome and Conversion of Phytapentaenal to 4,5-Didehydrogeranyl Geranoic Acid in Pig Liver Homogenate

  • Kim, Seon-Jae
    • Preventive Nutrition and Food Science
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    • v.5 no.4
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    • pp.234-238
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    • 2000
  • The cleavage products formed by autoxidation of phytofluene were evaluated in order to elucidate possible oxidation products of phytofluene under oxidative conditions. Phytofluene solubilized at 50$\mu$M in liposomal suspension was oxidized by incubating at 37$^{\circ}C$ for 72 h. Among a number of oxidation products formed, five products in the carbonyl compound fraction were identified as 6, 10, 14-trimethylpentadeca-3,5,9,13-tetraen-2-one, phytapentaenal, 5,9,13,17-tetramethyloctadeca-2,4,6,8,12,16-hexaenal, 5,9,13,17-tetramethyloctadeca-2,4,8,12,16-pentaenal, 2,7,11,15,19-pentamethylicosa-2,4,6,10,14,18-hexaenal and 4,9,13,17,21-pentamethyldocosa-2,4,6,8,12,16,20-heptaenal. These correspond to a series of products formed by cleavage in the respective eight conjugated double bonds of phytofluene. Also, 4,5-didehydorgeranyl geranoic acid was formed by autoxidation of phytofluene in liposomal suspension. The pig liver homogenate had the ability to convert phytapentaenal to 4,5-didehydrogeranyl geranoic acid, comparable to the conversion of all-trans-retinal to all-trans-retinoic acid. These results suggest that phytofluene is cleaved to a series of long-chain and short-chain carbonyl compounds under the oxidative condition in vitro and that phytapentaenal is further enzymatically converted to 4,5-didehydrogeranyl geranoic acid.

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In Vitro Selection of Hammerhead Ribozymes with Optimized Stems I and III

  • Sim, So-Yeong;Kim, Se-Mi;Kim, Ha-Dong;Ahn, Jeong-Keun;Lee, Young-Hoon;Cho, Bong-Rae;Park, In-Won
    • BMB Reports
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    • v.31 no.2
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    • pp.177-182
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    • 1998
  • A pool of cis-acting hammerhead ribozymes randomized in their substrate recognition sequences was constructed. A variety of active cis-acting ribozymes which had various structures of stems I and III was selected from the pool by in vitro selection. The selected ribozymes were cloned and sequenced. The relationship between the cleavage efficiency and base-pairing in stems I and III of the selected ribozymes was investigated. The ribozymes with the smaller difference in folding energies between the active conformation and the stable but inactive conformation showed a tendency to have the better cleavage efficiency. The optimum length of stem I was 5 or 6 bases while the longer stem III, in general, appeared to be required for efficient cleavage. The specificity of the ribozyme reaction is discussed in terms of the length of stems I and III.

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