• Title/Summary/Keyword: ribosome

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Analysis of Antifungal Proteins in Streptomyces sp. SAR01 (Streptomyces sp. SAR01 균주에서의 항진균 관련 단백질 분석)

  • 이영근;김재성;조규성;장병일;추철형
    • Korean Journal of Environmental Biology
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    • v.20 no.3
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    • pp.237-244
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    • 2002
  • To analyze proteins related to antifungal activity, SAR01 strain was isolated from seaweed and identified as Streptomyces sp. from the result of FAME (fatty acid methyl ester) analysis. The isolated strain had antifungal activities against T species of plant pathogenic fungi. Antifungal activity deficient mutant (SAR 535) of Streptomyces sp. SAR01 was induced by gamma radiation $(^{60}Co,\;5kGy)$. By 2 D electrophoresis analysis, 6 protein spots were found in wild strain (SAR01) but these spots disappeared in mutant strain (SAR535). Among them, 5 proteins showed similarities to heat shock protein 70(HSP70), Fe-containing superoxide dismutase II (Fe- SODII), ribosome recycling factor (RRF), 10 kDa chnperonin (GroES) and inorganic pyrophosphatase (PPAse), respectively. It suggested that the above 6 proteins could be closely related to the antifungal activity of Streptomyces sp. SAR01.

Effects of Chitosanoligosaccharide on the Mouse Hepatotoxicity Induced by Cadmium (카드뮴으로 유발된 생쥐 간독성에 대한 키토산올리고당의 효과)

  • Yoon, Jung-Sik;Roh, Young-Bok
    • Applied Microscopy
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    • v.32 no.4
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    • pp.361-376
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    • 2002
  • This research was conducted to determine the effects of chitosanoligosaccharide on liver poisoning induced by cadmium (Cd). Three groups of mice were used in this research. The group was only injected with cadmium (5.0 mg/kg; i.p.) (group Cd) and the other group was injected with cadmium and chitosanoligosaccharide (0.5% solution) at the same time (group Cd+Chi). In order to investigate the inhibitory action of chitosanoligosaccharide on liver damage, cadmium concentration in liver tissues and metallothionein (MT) concentration were relatively measured. In addition, histological observations were made to determine the morphologic injury of liver tissues. Cadmium concentration in liver tissues was drastically lower in groups Cd+Chi than in group Cd. MT concentration in liver tissues was lower in group Cd than in groups Cd+Chi. As the result of electron microscopic observation, mitochondria in group Cd showed a severe swelling phenomenon, RER fragment and ribosome dropout. However, in groups Cd+Chi, mitochondria with high electron density were distributed and RER forming a typical lamellae with ribosome was observed. From these results, cadmium toxicity on rat liver tissues could be lessened by chitosanoligosaccharide.

Cytological Changes Associated with the Exposure of Escherichia coli to Several Disinfectants: An Ultrastructural Study (수종(數種) 항균제(抗菌劑) 처리(處理)에 의(依)한 Escherichia coli의 미세구조(微細構造) 변화(變化))

  • Deung, Young-Kun;Koh, Choon-Myung;Kim, Sung-Kwang
    • The Journal of the Korean Society for Microbiology
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    • v.11 no.1
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    • pp.1-11
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    • 1976
  • An electron microscopic study has been made of the effects of change of cell structure of Escherichia coli treated with several disinfectants. The alterations observed as follows: 1. Nucleoid, cytoplasm which contain ribosomes and cell wall appeared to be composed of a parallel triple. layered membrane can observed in control Escherichia coli. 2. The nuclear material was no longer demonstrable in its normal sites. The cytoplasm lost its granularity, became homogeneous and disruption of cell wall were observed by the treatment with 70% ethyl alcohol and 3% $H_2O_2$. 3. Aggregation of ribosomes and condensation of nuclear material were also observed by the treatment of 5% lysol solution and 1% dodecyl-diamino-ethyl-glycin-hydrochloride. 4. In autoclaved group, the each layer of cell wall was separated and destroyed in some sites where cytoplasm was extracted.

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Inhibition of Hepatitis C Virus (HCV) Replication by Hammerhead Ribozyme Which Activity Can Be Allosterically Regulated by HCV NS5B RNA Replicase (C형 간염바이러스(HCV)의 NS5B RNA Replicase에 의해 활성이 유도되는 Hammerhead 리보자임에 의한 HCV 복제 억제 연구)

  • Lee, Chang-Ho;Lee, Seong-Wook
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.188-193
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    • 2011
  • As a specific and effective therapeutic genetic material against hepatitis C virus (HCV) multiplication, HCV internal ribosome entry site (IRES)-targeting hammerhead ribozyme which activity is allosterically regulated by HCV regulatory protein, NS5B RNA replicase, was constructed. The allosteric ribozyme was composed of sequence of RNA aptamer to HCV NS5B, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding NS5B to the aptamer, and sequence of ribozyme targeting +382 nucleotide of HCV IRES. With real-time PCR analysis, the ribozyme was found to efficiently inhibit HCV replicon replication in cells. Of note, the allosteric ribozyme was shown to inhibit HCV replicon replication more efficiently than either HCV genome-targeting ribozyme or NS5B aptamer only. This allosteric ribozyme can be used as a lead genetic agent for the specific and effective suppression of HCV replication.

Distribution and Ultrastructure of Dopaminergic Neurons in the Substantia Nigra of Mongolian Gerbil (Meriones unguiculates) (모래쥐 흑색질의 도파민성 신경세포의 분포와 미세구조)

  • Choi, Wol-Bong;Yoon, Sang-Seon;Ko, Byoung-Moon;Jo, Seung-Mook;Nam, Seong-Ahn;Choi, Chang-Do
    • Applied Microscopy
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    • v.27 no.4
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    • pp.461-472
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    • 1997
  • The substantia nigra of the Mongolian gerbil was studies by tyrosine hydroxylase immunohistochemistry and immunoelectron microscopy with preembedding method. The purpose was to obtain information on the distribution and ultrastructure of the Tyrosine hydroxylase immunoreactive and dopaminergic neurons in the substantia nigra, in order to provide the necessary background for the gerbil. Large number of tyrosine hydroxylase immunoreactive neurons were located in the compact part of substantia nigra. Findings in the gerbil, compared to observations in the other species, included the presence of prominent bundles of tyrosine hydroxylase immunoreactive cytoplasmic processes passing in the dorsoventral direction from pars compacta into pars reticulata at middle and caudal levels of the substantia nigra, and the presence of a distinct tyrosine hydroxylase immunoreactive substantia nigra pars lateralis. Tyrosine hydroxylase immunoreactive neurons had well-developed cell organelles, especially rough endoplasmic reticulum, free ribosome and poly-ribosome, and showed the infoldings of the nuclear envelope. We anticipate that the present description of the cellular organization of the tyrosine hydroxylase immunoreactive dopaminergic area in the substantia nigra of gerbil will be useful for the animal experimental model of Parkinson's disease.

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Functional Analysis of the Invariant Residue G791 of Escherichia coli 16S rRNA

  • Song, Woo-Seok;Kim, Hong-Man;Kim, Jae-Hong;Sim, Se-Hoon;Ryou, Sang-Mi;Kim, Sang-Goo;Cha, Chang-Jun;Cunningham, Philip R.;Bae, Jee-Hyeon;Lee, Kang-Seok
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.418-421
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    • 2007
  • The nucleotide at position 791(G791) of E. coli 16S rRNA was previously identified as an invariant residue for ribosomal function. In order to characterize the functional role of G791, base substitutions were introduced at this position, and mutant ribosomes were analyzed with regard to their protein synthesis ability, via the use of a specialized ribosome system. These ribosomal RNA mutations attenuated the ability of ribosomes to conduct protein synthesis by more than 65%. A transition mutation (G to A) exerted a moderate effect on ribosomal function, whereas a transversion mutation (G to C or U) resulted in a loss of protein synthesis ability of more than 90%. The sucrose gradient profiles of ribosomes and primer extension analysis showed that the loss of protein-synthesis ability of mutant ribosomes harboring a base substitution from G to U at position 791 stems partially from its inability to form 70S ribosomes. These findings show the involvement of the nucleotide at position 791 in the association of ribosomal subunits and protein synthesis steps after 70S formation, as well as the possibility of using 16S rRNA mutated at position 791 for the selection of second-site revertants in order to identify ligands that interact with G791 in protein synthesis.

Functional Analysis of the Residue 789 in Escherichia coli 16S rRNA and Development of a Method to Select Second-site Revertants (Escherichia coli 16S rRNA의 789 염기의 기능분석 및 이차복귀돌연변이체 발췌를 위한 방법 개발)

  • Kim Jong-Myung;Go Ha-Young;Song Woo-Seok;Ryou Sang-Mi;Lee Kang-Seok
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.156-159
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    • 2006
  • A base substitution was introduced at the position 789 in Escherichia coli 16S rRNA, which was previously identified as an invariant residue for ribosome function and the ability of the mutant ribosomes to translate chloramphenicol acetyltransfernse mRNA was measured by determining the degree of resistance to chloramphenicol of cells expressing these mutant ribosomes. As expected, mutant ribosomes containing a base sub-stitution at the position 789 showed significantly reduced protein-synthesis ability and to identify a functional role played by this residue in protein synthesis, we developed an efficient genetic method to select second-site revertants in 16S rRNA that restore protein-synthesis function to these mutant ribosomes.

Combinatorial Fine-Tuning of Phospholipase D Expression by Bacillus subtilis WB600 for the Production of Phosphatidylserine

  • Huang, Tingting;Lv, Xueqin;Li, Jianghua;Shin, Hyun-dong;Du, Guocheng;Liu, Long
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2046-2056
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    • 2018
  • Phospholipase D has great commercial value due to its transphosphatidylation products that can be used in the food and medicine industries. In order to construct a strain for use in the production of PLD, we employed a series of combinatorial strategies to increase PLD expression in Bacillus subtilis WB600. These strategies included screening of signal peptides, selection of different plasmids, and optimization of the sequences of the ribosome-binding site (RBS) and the spacer region. We found that using the signal peptide amyE results in the highest extracellular PLD activity (11.3 U/ml) and in a PLD expression level 5.27-fold higher than when the endogenous signal peptide is used. Furthermore, the strain harboring the recombinant expression plasmid pMA0911-PLD-amyE-his produced PLD with activity enhanced by 69.03% (19.1 U/ml). We then used the online tool \RBS Calculator v2.0 to optimize the sequences of the RBS and the spacer. Using the optimized sequences resulted in an increase in the enzyme activity by about 26.7% (24.2 U/ml). In addition, we found through a transfer experiment that the retention rate of the recombinant plasmid after 5 generations was still 100%. The final product, phosphatidylserine (PS), was successfully detected, with transphosphatidylation selectivity at 74.6%. This is similar to the values for the original producer.

Transcriptome-wide analysis reveals gluten-induced suppression of small intestine development in young chickens

  • Darae, Kang;Donghyun, Shin;Hosung, Choe;Doyon, Hwang;Andrew Wange, Bugenyi;Chong-Sam, Na;Hak-Kyo, Lee;Jaeyoung, Heo;Kwanseob, Shim
    • Journal of Animal Science and Technology
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    • v.64 no.4
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    • pp.752-769
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    • 2022
  • Wheat gluten is an increasingly common ingredient in poultry diets but its impact on the small intestine in chicken is not fully understood. This study aimed to identify effects of high-gluten diets on chicken small intestines and the variation of their associated transcriptional responses by age. A total of 120 broilers (Ross Strain) were used to perform two animal experiments consisting of two gluten inclusion levels (0% or 25%) by bird's age (1 week or 4 weeks). Transcriptomics and histochemical techniques were employed to study the effect of gluten on their duodenal mucosa using randomly selected 12 broilers (3 chicks per group). A reduction in feed intake and body weight gain was found in the broilers fed a high-gluten containing diet at both ages. Histochemical photomicrographs showed a reduced villus height to crypt depth ratio in the duodenum of gluten-fed broilers at 1 week. We found mainly a significant effect on the gene expression of duodenal mucosa in gluten-fed broilers at 1 week (289 differentially expressed genes [DEGs]). Pathway analyses revealed that the significant DEGs were mainly involved in ribosome, oxidative phosphorylation, and peroxisome proliferator-activated receptor (PPAR) signaling pathways. These pathways are involved in ribosome protein biogenesis, oxidative phosphorylation and fatty acid metabolism, respectively. Our results suggest a pattern of differential gene expression in these pathways that can be linked to chronic inflammation, suppression of cell proliferation, cell cycle arrest and apoptosis. And via such a mode of action, high-gluten inclusion levels in poultry diets could lead to the observed retardation of villi development in the duodenal mucosa of young broiler chicken.

SAMD13 as a Novel Prognostic Biomarker and its Correlation with Infiltrating Immune Cells in Hepatocellular Carcinoma

  • Hye-Ran Kim;Choong Won Seo;Jae-Ho Lee;Sang Jun Han;Jongwan Kim
    • Biomedical Science Letters
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    • v.28 no.4
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    • pp.260-275
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    • 2022
  • Sterile alpha motif (SAM) domains bind to various proteins, lipids, and RNAs. However, these domains have not yet been analyzed as prognostic biomarkers. In this study, SAM domain containing 13 (SAMD13), a member of the SAM domain, was evaluated to identify a novel prognostic biomarker in various human cancers, including hepatocellular carcinoma (HCC). Moreover, we identified a correlation between SAMD13 expression and immune cell infiltration in HCC. We performed bioinformatics analysis using online databases, such as Tumor Immune Estimation Resource, UALCAN, Kaplan-Meier plotter, LinkedOmics, and Gene Expression Profiling Interactive Analysis2. SAMD13 expression in HCC samples was significantly higher than that in normal liver tissue; additionally, SAMD13 was higher in primary tumors, various stages of cancer and grades of tumor, and status of nodal metastasis. Higher SAMD13 expression was also associated with poorer prognosis. SAMD13 expression positively correlated with CD8+ T cells, CD4+ T cells, B cells, neutrophils, macrophages, and dendritic cells. In the analysis of SAMD13 co-expression networks, positively related genes of SAMD13 were associated with a high hazard ratio in different types of cancer, including HCC. In biological function of SAMD13, SAMD13 mainly include spliceosome, ribosome biogenesis in eukaryote, ribosome, etc. These results suggest that SAMD13 may serve as a novel prognostic biomarker for HCC diagnosis and provide novel insights into tumor immunology in HCC.