• Title/Summary/Keyword: Ribosome

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Cloning and Sequence Analysis of the trpB, trpA and 3' trpC(F) Gens of Vibrio metschnikovii Strain RH530 (Vibrio metschnikovii 균주 RH530의 trpB, trpA 그리고 3' trpC(F) 유전자의 클로닝 및 염기서열 결정)

  • Kwon, Yong-Tae;Kim, Jin-Oh;Yoo, Young-Dong;Rho, Hyune-Mo
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.120-125
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    • 1994
  • The genes, trpB, trpA and 3’ trpC(F) of Vibrio metschnikovii strain RH530 were cloned and sequenced. The trpB and trpA genes had open reading frames of 1,173 bp and 804 bp encoding 391 and 268 amino acids, respectively. The trpB and trpA genes had conventional ribosome-binding sequences and overlapped with each other by one nucleotide, suggesting that these two genes are translationally coupled. 115 nucleotide upstream the trpB start codon, tjere was an incomplete open reading frame of the 3’-end of the trpC(F). The amino acid sequences of trpB, trpA and trpC(F) of V. metschnikovii RH530 had identities of 64.2%, 82.4% and 73.7% respectively, for those of V. parahaemolyticus; 58.7%, 72.3% and 54.9%, respectively, for Salmonella typhimurium; and 42.6%. 54.1% and 12.5%, respectively, for brevibacterium lactofermentum. The genetic organization of these genes, especially in the noncoding region between trpC(F) and trpB, was distinct from that of Enterobacteriaceae.

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Increased Production of an Alkaline Protease from Bacillus clausii I-52 by Chromosomal Integration (Bacillus clausii I-52의 Chromosomal Integration에 의한 Alkaline Protease의 생산성 향상)

  • Joo, Han-Seung;Park, Dong-Chul;Choi, Jang-Won
    • Journal of agriculture & life science
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    • v.46 no.1
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    • pp.163-176
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    • 2012
  • TTo increase productivity of a strong extracellular alkaline protease (BCAP), stable strains of Bacillus clausii I-52 carrying another copy of BCAP gene in the chromosome were developed. Integrative vector, pHPS9-fuBCAP carrying BCAP promoter, ribosome binding site, signal sequence and active protease gene was constructed and transferred into B. clausii I-52, and integration of the constructed plasmid into chromosome was identified by PCR. An investigation was carried out on BCAP production by B. clausii I-52 and transformant C5 showing the highest relative activity of alkaline protease using submerged fermentation. Maximum enzyme activity was produced when cells were grown under the submerged fermentation conditions at $37^{\circ}C$ for 48 h with an aeration rate of 1 vvm and agitation rate of 650 rpm in a optimized medium (soybean meal 2%, wheat flour 1%, sodium citrate 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_47H_2O$ 0.01%, $FeSO_47H_2O$ 0.05%, liquid maltose 2.5%, $Na_2CO_3$ 0.6%). A protease yield of approximately 134,670U/ml was achieved using an optimized media, which show an increase of approximately 1.6-fold compared to that of non-transformant (83,960 U/ml). When the stability of transformant C5 was examined, the integrated plasmid pHPS9-fuBCAP was detected in the transformant after cultivation for 8 days, suggesting that it maintained stably in the chromosomal DNA of transformant C5.

Structural Disorganization of Intestinal Tumor Spheroid by Microbial Ribotoxins (방사선 모사 미생물 유래 리보솜 스트레스에 의한 대장암 스페로이드 구조 결함 유발)

  • Kim, Juil;Kim, Joongkon;Yu, Mira;Moon, Yuseok
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.164-171
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    • 2019
  • Radiation therapy has many side effects, such as digestive mucosal ulcers, without regard to its efficacy. The purpose of this study is to address an alternative method to replace the limitation of radiation therapy using radiomimetic microbial ribotoxins. In the evaluation of cancer therapy, we analyzed the formation of colorectal cancer (CRC) cell spheroids, which can take into account the heterogeneous cellular constitution, tumor stem cells, and the surrounding microenvironment. Ribotoxic stress interfered with the spheroid structure composed of relatively small clusters. Spheroids under ribotoxic stress were structurally sparse and their shrinkage was very slow. In the control group, the clusters of strongly aggregated cells were resistant to physical stress, but the ribotoxic stress-exposed spheroids were easily broken up by the physical stress. Moreover, the ribosome-insulted CRC cells slowly migrated to form clusters and the cell-cell junctional points in the ribosome-insulted spheroids were rarer than those in the control CRC spheroid. Moreover, levels of the cell-to-cell junctional protein E-cadherin were suppressed by ribotoxic stress in both allograft and xenograft spheroids. In conclusion, the radiomimetic microbial ribotoxins induced structural defects in CRC cell spheroids via retardation of migration and cell-cell junction in the formation of three-dimensional structures, and provides a basis for the mechanism of pharmacological radiomimetic anticancer actions as an alternate to radiotherapy against cancer.

Multiple functions of human UV DNA repair endonuclease III

  • Jang, Chang-Young;Kim, Joon
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.182-185
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    • 2002
  • There are 3 UV DNA repair endonuclease activities in mammalian cells that cleave UV -irradiated DNA. Interestingly, mammalian UV endonuclease III with MW of 26.7kD has a lyase activity on AP sites. It also cleaves the phosphodiester bond within a cyclobutane pyrimidine dimer. Genomic analysis of human repair endonuclease III gene revealed that this gene has 100% sequence identity with ribosomal protein S3 (rpS3). Therefore, rpS3 seems to function both in translation and in DNA repair. This gene of about 6.1 kb contains 6 introns and 7 exons, and the first and fifth introns of human rpS3 gene contain functional U15 small nucleolar (sno) RNAs which appear to be involved in ribosome assembly. It is to be noted that the column profile of the endonuclease activity of rpS3 appears to be altered in Xeroderma Pigmentosum (XP) group D cells compared to normal cells indicating that this protein is involved in XP disease as well. XP is a human disease characterized by high sensitivity of skin by UV- or sun-light irradiation and by high frequency of developing skin cancers. We also report here that rpS3 protein is involved in other cellular functions.

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Ultrastructure of Cryptococcus neoformans in the Skin Tissue (조직내 Cryptococcus neoformans의 전자현미경적 관찰)

  • Seo, Young-Hoon;Kwon, Tae-Jung;Kim, Chung-Sook
    • Applied Microscopy
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    • v.12 no.1
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    • pp.49-56
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    • 1982
  • A case of systemic cryptococcosis developed in 4 year old boy was described and illustrated by light and electron microscope. Light microscopically, the upper dermis of the skin showed chronic nonspecific inflammation with numerous spherical spores surrounded by a clear halo created by the wide gelatinous capsule. Ultrastructurally, the C. neoformans showed the wide capsule containing microfibrils that appeared to radiate from the cell wall and to coil and interwine in various directions. The cell was uninucleate with a single nucleolus. Along the inner nuclear envelope, numerous small vesicles were present. In addition, C. neoformans presented membranous organelles derived from the plasma membrane and comparable to bacterial mesosomes.

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Ultrastructure of Haustorial Cells of Cuscuta australis R. Brown (실새삼(Cuscuta australis R. Brown) 흡기세포(吸器細胞)의 미세구조(微細構造))

  • Lee, Chai-Doo;Lee, Kyu-Bae
    • Applied Microscopy
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    • v.16 no.2
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    • pp.49-60
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    • 1986
  • Ultrastructures of the large and elongated cells (idioblasts) in the haustorium of a parasitic angiosperm, Cuscuta australis R. Brown growing on the host plant, Trifolium repens L. were investigated by the electron microscopy. The idioblasts were characterized by the presence of a large nucleus, small vacuoles, and dense cytoplasm including a number of various cell organelles such as ribosome, rough endoplasmic reticulum(r-ER), mitochondrion, dictyosome, proplastid, multilamellar structure(MLS), microfilament bundle(MFB), and cytosegresome. Therefore, it is suggested that the idioblasts are metabolical1y very active. Particularly, MLS, MFB, and cytosegresome observed in this study did not appear in the haustorial cells of the other parasitic angiosperms. MLS was transformed into vacuole and also incorporated with cell wall. MFB composed of microfilaments, about each 7.5 nm in diameter, was observed in nucleus and also cytoplasm. Some types of MFB were distinguished on the basis of arrangement of microfilaments. A part of cytoplasm sequestered by stacked cisternae of smooth ER(s-ER), cytosegresome, was altered into a vacuole which was formed by digestion of the sequestered cytoplasm and of cisternae of s-ER. Cell organelles such as MLS, MFB, and cytosegresome were discussed in relation to the metabolic control of the idioblasts.

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Current insights into inherited bone marrow failure syndromes

  • Chung, Nack-Gyun;Kim, Myungshin
    • Clinical and Experimental Pediatrics
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    • v.57 no.8
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    • pp.337-344
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    • 2014
  • Inherited bone marrow failure syndrome (IBMFS) encompasses a heterogeneous and complex group of genetic disorders characterized by physical malformations, insufficient blood cell production, and increased risk of malignancies. They often have substantial phenotype overlap, and therefore, genotyping is often a critical means of establishing a diagnosis. Current advances in the field of IBMFSs have identified multiple genes associated with IBMFSs and their pathways: genes involved in ribosome biogenesis, such as those associated with Diamond-Blackfan anemia and Shwachman-Diamond syndrome; genes involved in telomere maintenance, such as dyskeratosis congenita genes; genes encoding neutrophil elastase or neutrophil adhesion and mobility associated with severe congenital neutropenia; and genes involved in DNA recombination repair, such as those associated with Fanconi anemia. Early and adequate genetic diagnosis is required for proper management and follow-up in clinical practice. Recent advances using new molecular technologies, including next generation sequencing (NGS), have helped identify new candidate genes associated with the development of bone marrow failure. Targeted NGS using panels of large numbers of genes is rapidly gaining potential for use as a cost-effective diagnostic tool for the identification of mutations in newly diagnosed patients. In this review, we have described recent insights into IBMFS and how they are advancing our understanding of the disease's pathophysiology; we have also discussed the possible implications they will have in clinical practice for Korean patients.

Two distinct nodes of translational inhibition in the Integrated Stress Response

  • Ryoo, Hyung Don;Vasudevan, Deepika
    • BMB Reports
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    • v.50 no.11
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    • pp.539-545
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    • 2017
  • The Integrated Stress Response (ISR) refers to a signaling pathway initiated by stress-activated $eIF2{\alpha}$ kinases. Once activated, the pathway causes attenuation of global mRNA translation while also paradoxically inducing stress response gene expression. A detailed analysis of this pathway has helped us better understand how stressed cells coordinate gene expression at translational and transcriptional levels. The translational attenuation associated with this pathway has been largely attributed to the phosphorylation of the translational initiation factor $eIF2{\alpha}$. However, independent studies are now pointing to a second translational regulation step involving a downstream ISR target, 4E-BP, in the inhibition of eIF4E and specifically cap-dependent translation. The activation of 4E-BP is consistent with previous reports implicating the roles of 4E-BP resistant, Internal Ribosome Entry Site (IRES) dependent translation in ISR active cells. In this review, we provide an overview of the translation inhibition mechanisms engaged by the ISR and how they impact the translation of stress response genes.

Local protein synthesis in neuronal axons: why and how we study

  • Kim, Eunjin;Jung, Hosung
    • BMB Reports
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    • v.48 no.3
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    • pp.139-146
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    • 2015
  • Adaptive brain function and synaptic plasticity rely on dynamic regulation of local proteome. One way for the neuron to introduce new proteins to the axon terminal is to transport those from the cell body, which had long been thought as the only source of axonal proteins. Another way, which is the topic of this review, is synthesizing proteins on site by local mRNA translation. Recent evidence indicates that the axon stores a reservoir of translationally silent mRNAs and regulates their expression solely by translational control. Different stimuli to axons, such as guidance cues, growth factors, and nerve injury, promote translation of selective mRNAs, a process required for the axon's ability to respond to these cues. One of the critical questions in the field of axonal protein synthesis is how mRNA-specific local translation is regulated by extracellular cues. Here, we review current experimental techniques that can be used to answer this question. Furthermore, we discuss how new technologies can help us understand what biological processes are regulated by axonal protein synthesis in vivo.

Cell-type-specific Gene Expression Patterns in Human Carcinoma Cells followed by Irradiation (방사선에 의한 암세포주 특이적 유전자 발현 양상)

  • Park Ji-Yoon;Kim Jin-Kyu;Chai Young Gyu
    • Korean Journal of Environmental Biology
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    • v.23 no.2 s.58
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    • pp.152-156
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    • 2005
  • Ionizing radiation is a well- known therapy factor for human carcinoma cells. Genotoxic stress mediates cell cycle control, transcription and cellular signaling. In this work, we have used a microarray hybridization approach to characterize the cell type-specific transcriptional response of human carcinoma MCF-7 and HeLa cell line to $\gamma-radiation$, such as 4Gy 4hr. We found that exposure to $\gamma-ray$ alters by at least a $log_2$ factor of 1.0 the expression of known genes. Of the 27 genes affected by irradiation, 11 are down- regulated in MCF-7 cells and 2 genes induced by radiation,15 are repressed in HeLa cells. Many genes were involved in known damage- response pathways for cell cycling, transcription factor and cellular signaling response. However, in MCF-7 cells, we observed gene expression pattern in chromatin, apoptosis, stress, differentiation, cytokine, metabolism, ribosome and calcium. In HeLa cells, it showed clearly the expression changes in adhesion and migration, lysosome, brain, genome instability and translation. These insights reveal new therapy directions for studying the human carcinoma cell response to radiation.