• 제목/요약/키워드: ribosomal protein S3

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MCF-7 세포주에서$\gamma$선에 의한 세포신호 전달 관련 유전자의 발현 양상의 분석 (Signal Transduction-related Gene Expression Analysis in MCF-7 followed by $\gamma$-radiation)

  • 박지윤;황창일;박웅양;김진규;채영규
    • 환경생물
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    • 제21권1호
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    • pp.52-55
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    • 2003
  • There is considerable evidence that ionizing radiation (IR) mediates checkpoint control, repair and cell death. In this study, we have used a high density microarray hybridization approach to characterize the transcriptional response of human breast carcinoma MCF-7 cell line to ${\gamma}$-radiation, such as 4 Gy 4 hr, 8 Gy 4 hr, and 8 Gy 12 hr. We found that exposure to ${\gamma}$-ray alters by at least a $log_2$ factor of 1.0 the expression of 115 known genes. Of the 66 genes affected by ${\gamma}$-radiation, 49 are down-regulated. In our results, the cellular response to irradiation includes induction of the c-jun and EGR1 early response genes. The present work has examined potential cytoplasmic signaling cascades that transduce IR-induced signals to the nucleus. 40S ribosomal protein s6 kinase modulates the activities of the mitogen activated protein kinase (MAPK) and c-Jun $NH_2$-terminal kinase (JNK1) cascades in human monocytic leukemia (U937/pREP4) cells. 14-3-3 family members are dimeric phosphoserine -binding proteins that participate in signal transduction and checkpoint control pathways.

Analysis of partial cDNA sequence from Theileria sergenti

  • Park, Jin-ho;Chae, Joon-seok;Kim, Dae-hyuk;Jang, Yong-suk;Kwon, Oh-deog;Lee, Joo-mook
    • 대한수의학회지
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    • 제39권4호
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    • pp.797-805
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    • 1999
  • T sergenti cDNA library were constructed to get a more broad information about the structural, functional or antigenic properties of the proteins, and analyzes for their partial cDNA sequences and expression sequences tags(ESTg). The mRNA were purified from T sergenti isolates to identify the information of antigen gene, then first and second strand cDNA was synthesized. EcoR I adaptor ligation and Xho I enzyme restriction were used to the synthesized cDNA, and ligated into a Uni-ZAP XR vector. T sergenti cDNA library was constructed with packaging and amplification in vitro. Antibody screening was performed with constructed T sergenti cDNA library using antisera against T sergenti. Among those clones, eight phagemids were rescued from the recombinant in vivo excision with f1 helper phage. Using the analysis of endonuclease restriction and PCR, the recombinant cDNA were proved having a 0.5-3.0kb of inserts. The eight of partial cDNA clones' sequences were obtained and examined for their homology using BLASTN and BLASTX. The eight of sequenced clones were classified into three groups according to the basis of database searches. A total 3,045bp of partial cDNA sequence were determined from six clones. The putatively identified clones contain a cytochrome c gene, a heat shock protein gene, a cyclophilin gene, and a ribosomal protein gene. The unidentified clones have a homology to ATP-binding protein(mtrA) gene of S argillaceus, DNA-binding protein(DBP) gene of Pseudorabies virus 85kDa merozoite protein gene of B bovis, mRNA spm1 protein of T annulata and glycine-rich RNA-binding protein mRNA of O sativa etc.

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Biosynthesis of 3-Hydroxy-5-Methyl-O-Methyltyrosine in the Saframycin/Safracin Biosynthetic Pathway

  • Fu, Cheng-Yu;Tang, Man-Cheng;Peng, Chao;Li, Lei;He, Yan-Ling;Liu, Wen;Tang, Gong-Li
    • Journal of Microbiology and Biotechnology
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    • 제19권5호
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    • pp.439-446
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    • 2009
  • The biosynthesis study of antibiotics saframycin (SFM) in Streptomyces lavendulae and safracin (SAC) in Pseudomonas fluorescens demonstrated that 3-hydroxy-S-methyl-O-methyltyrosine (3hSmOmTyr), a nonproteinogenic amino acid, is the precursor of the tetrahydroisoquinoline molecular core. In the biosynthetic gene cluster of SAC/SFM, sacD/sfmD encodes a protein with high homology to each other but no sequence similarity to other known enzymes; sacF/sfmM2 and sacG/sfmM3 encode methyltransferases for C-methylation and O-methylation; and sacE/sfinF encodes a small protein with significant sequence similarity to the MbtH-like proteins, which are frequently found in the biosynthetic pathways of non ribosomal peptide antibiotics and siderophores. To address their function, the biosynthetic cassette of 3h5mOmTyr was heterologously expressed in S. coelicolor and P. putida, and an in-frame deletion and complementation in trans were carried out. The results revealed that (i) SfmD catalyzes the hydroxylation of aromatic rings; (ii) sacD/sacF/sacG in the SAC gene cluster and sfmD/sfmM2/sfmM3 in the SFM cluster are sufficient for the biosynthesis of 3h5mOmTyr; and (iii) the mbtH-like gene is not required for the biosynthesis of the 3h5mOmTyr precursor.

자가영양과 타가영양 조건하에서 Thiobacillus novellus의 생리 및 성장 (Growth and Physiology of Thiobacillus novellus under Autotrophic and Heterotrophic Conditions)

  • 박인국
    • 미생물학회지
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    • 제29권4호
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    • pp.263-266
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    • 1991
  • The growth of T. novellus in auto trophic and geterotrophic media was studied to determine the time required for cells to enter stationary phase and relative percentage of ribosomal proteins. When T. novellus was grown autotrophically, growth proceeded at a slow rate characteristic of autotrophs and did not enter log phase until the end of the first day. Logarithmic growth proceeded for 3-4 days at which time the cells entered the stationary phase. In particular, logarithmic growth was accompanied by decreasing pH of culture media and in the stationary phase the pH levelled off at 6.0, a decrease of 1.6 pH value compared to original pH of media. The pH decrease was greatest during log phase when cells oxidized thiosulfate to $H_{2}$$SO_{4}$. The doubling time was about 26h. In heterotrophic media growth proceeded at a much faster rate and cells entered stationary phase 20-22h after inoculation. The doubling time was 3h. The protein content of the ribosomes in T. novellus grown heterotrophically was 4.2% greater than those from the organism grown autotrophically.

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Comparative Analyses of Tomato yellow leaf curl virus C4 Protein-Interacting Host Proteins in Healthy and Infected Tomato Tissues

  • Kim, Namgyu;Kim, Jinnyun;Bang, Bongjun;Kim, Inyoung;Lee, Hyun-Hee;Park, Jungwook;Seo, Young-Su
    • The Plant Pathology Journal
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    • 제32권5호
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    • pp.377-387
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    • 2016
  • Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus, is one of the most important viruses of cultivated tomatoes worldwide, mainly causing yellowing and curling of leaves with stunting in plants. TYLCV causes severe problems in sub-tropical and tropical countries, as well as in Korea. However, the mechanism of TYLCV infection remains unclear, although the function of each viral component has been identified. TYLCV C4 codes for a small protein involved in various cellular functions, including symptom determination, gene silencing, viral movement, and induction of the plant defense response. In this study, through yeast-two hybrid screenings, we identified TYLCV C4-interacting host proteins from both healthy and symptom-exhibiting tomato tissues, to determine the role of TYLCV C4 proteins in the infection processes. Comparative analyses of 28 proteins from healthy tissues and 36 from infected tissues showing interactions with TYLCV C4 indicated that TYLCV C4 mainly interacts with host proteins involved in translation, ubiquitination, and plant defense, and most interacting proteins differed between the two tissues but belong to similar molecular functional categories. Four proteins-two ribosomal proteins, S-adenosyl-L-homocysteine hydrolase, and 14-3-3 family protein-were detected in both tissues. Furthermore, the identified proteins in symptom-exhibiting tissues showed greater involvement in plant defenses. Some are key regulators, such as receptor-like kinases and pathogenesis-related proteins, of plant defenses. Thus, TYLCV C4 may contribute to the suppression of host defense during TYLCV infection and be involved in ubiquitination for viral infection.

폐암 세포주에서 광역학 치료에 의한 유전자 발현 분석 (Gene Expression Profile of Lung Cancer Cells Following Photodynamic Therapy)

  • 성지현;이미은;한선숙;이승준;하권수;김우진
    • Tuberculosis and Respiratory Diseases
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    • 제63권1호
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    • pp.52-58
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    • 2007
  • 연구배경: 광역학 치료는 폐암 치료에 실질적으로 이용 가능하며, 많은 연구들에서 폐암 세포에서 세포사멸을 일으킨다는 것이 이미 알려져 있다. 그러나 이 세포사멸의 기전은 아직 정확히 알려져 있지 않으며, 이에 암세포의 전사에서 초기 변화가 어떻게 일어나는 지를 알아보기 위하여 실험을 수행하였다. 방 법: 광과민성 물질인 DH-I-180-3으로 A549 세포에 처리를 하고 광역학 치료를 한 후 관찰하였다. 광역학 치료 후 DEG kit를 이용하여 폐암 세포주에서의 유전자 발현을 보았으며, 유세포 분석기를 이용하여 세포 사멸을 측정하였다. 광역학 치료 후 의미있는 변화를 보인 유전자는 염기서열분석으로 확인하였다. 결 과: 유세포분석 결과 폐암세포주는 대부분 세포괴사에 의하여 사멸되었다.광역학 치료 후, 9개의 유전자에서 명확한 변화가 있음을 발견했으며 이 중8개의 유전자를 밝혀내었다. 3-phosphoglycerate dehydrogenase와 리보솜 단백질 S29의 유전자 발현이 증가되어 있었으며, carbonic anhydrase XII, clusterin, MRP3s1 protein, complement 3, membrane cofactor protein, ${\beta}$-1 integrin의 유전자 발현은 감소되어 있었다. 결 론: 본 연구는 광과민성 물질인 DH-I-180-3을 이용한 광역학 치료에서 폐암 세포의 세포사멸의 주된 기전이 세포괴사에 의해 이루어 진 것임을 밝혀냈으며, 이와 관련된 유전자들 대부분이 막단백의 변화를 통해 이루어짐을 알 수 있었다.

Novel Anti-Angiogenic and Anti-Tumour Activities of the N-Terminal Domain of NOEY2 via Binding to VEGFR-2 in Ovarian Cancer

  • Rho, Seung Bae;Lee, Keun Woo;Lee, Seung-Hoon;Byun, Hyun Jung;Kim, Boh-Ram;Lee, Chang Hoon
    • Biomolecules & Therapeutics
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    • 제29권5호
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    • pp.506-518
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    • 2021
  • The imprinted tumour suppressor NOEY2 is downregulated in various cancer types, including ovarian cancers. Recent data suggest that NOEY2 plays an essential role in regulating the cell cycle, angiogenesis and autophagy in tumorigenesis. However, its detailed molecular function and mechanisms in ovarian tumours remain unclear. In this report, we initially demonstrated the inhibitory effect of NOEY2 on tumour growth by utilising a xenograft tumour model. NOEY2 attenuated the cell growth approximately fourfold and significantly reduced tumour vascularity. NOEY2 inhibited the phosphorylation of the signalling components downstream of phosphatidylinositol-3'-kinase (PI3K), including phosphoinositide-dependent protein kinase 1 (PDK-1), tuberous sclerosis complex 2 (TSC-2) and p70 ribosomal protein S6 kinase (p70S6K), during ovarian tumour progression via direct binding to vascular endothelial growth factor receptor-2 (VEGFR-2). Particularly, the N-terminal domain of NOEY2 (NOEY2-N) had a potent anti-angiogenic activity and dramatically downregulated VEGF and hypoxia-inducible factor-1α (HIF-1α), key regulators of angiogenesis. Since no X-ray or nuclear magnetic resonance structures is available for NOEY2, we constructed the three-dimensional structure of this protein via molecular modelling methods, such as homology modelling and molecular dynamic simulations. Thereby, Lys15 and Arg16 appeared as key residues in the N-terminal domain. We also found that NOEY2-N acts as a potent inhibitor of tumorigenesis and angiogenesis. These findings provide convincing evidence that NOEY2-N regulates endothelial cell function and angiogenesis by interrupting the VEGFR-2/PDK-1/GSK-3β signal transduction and thus strongly suggest that NOEY2-N might serve as a novel anti-tumour and anti-angiogenic agent against many diseases, including ovarian cancer.

토양 방선균인 Gordonia sp. MMS17-SY073 균주의 유전체 분석 (Complete genome sequence of Gordonia sp. MMS17-SY073, a soil actinobacterium)

  • 김영석;김승범
    • 미생물학회지
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    • 제55권3호
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    • pp.303-305
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    • 2019
  • 섬 해안가 토양에서 방선균주 Gordonia sp. MMS17-SY073를 분리하여 유전체 분석을 실시하였고, 그 결과 5,962,176 염기쌍 및 67.4%의 G + C 함량으로 이루어진 유전체 정보를 확보하였다. 유전정보 분석 결과 총 5,201개 단백질 지정 유전자, 6개 rRNA 유전자 및 45개 tRNA 유전자를 확인하였다. MMS17-SY073 균주는 16S rRNA 유전자를 이용한 분석 결과 분류학적으로 Gordonia soli의 표준균주와 가장 가까웠으며 그 유사도는 98.5%로 나타났다. MMS17-SY073 균주는 non-ribosomal peptide synthetase 유형을 비롯한 다수의 이차대사산물 생합성 유전자를 보유하고 있는 것으로 나타났다.

Effects of protein content and the inclusion of protein sources with different amino acid release dynamics on the nitrogen utilization of weaned piglets

  • Hu, Nianzhi;Shen, Zhiwen;Pan, Li;Qin, Guixin;Zhao, Yuan;Bao, Nan
    • Animal Bioscience
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    • 제35권2호
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    • pp.260-271
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    • 2022
  • Objective: We aimed to investigate the effect of the differing amino acid (AA) release dynamics of two protein sources on the growth performance, nitrogen deposition, plasma biochemical parameters, and muscle synthesis and degradation of piglets when included in their diets at normal and low concentrations. Methods: Forty-eight piglets (Duroc×Landrace×Large White) with initial body weight of 7.45±0.58 kg were assigned to six groups and fed one of 6 diets. The 6 dietary treatments were arranged by 3×2 factorial with 3 protein sources and 2 dietary protein levels. They are NCAS (a normal protein content with casein), NBlend (a normal protein content with blend of casein and corn gluten meal), NCGM (a normal protein content with corn gluten meal), LCAS (a low protein content with casein), LBlend (a low protein content with blend of casein and corn gluten meal), LCGM (a low protein content with corn gluten meal). The release dynamics of AA in these diets were determined by in vitro digestion. The digestibility, utilization and biological value of nitrogen in piglets were determined by micro Kjeldahl method. Plasma insulin was measured by enzyme-linked immunosorbent assay kits. The protein expression of mediators of muscle synthesis and degradation was determined by western blotting. Results: Although the consumption of a low-protein diet supplemented with crystalline AA was associated with greater nitrogen digestion and utilization (p<0.05), the final body weight, growth performance, nitrogen deposition, and phosphorylation of ribosomal protein S6 kinase 1 and eIF4E binding protein 1 in the muscle of pigs in the low-protein diet-fed groups were lower than those of the normal-protein diet-fed groups (p<0.05) because of the absence of non-essential AA. Because of the more balanced release of AA, the casein (CAS) and Blend-fed groups showed superior growth performance, final body weight and nitrogen deposition, and lower expression of muscle ring finger 1 and muscle atrophy F-box than the CGM-fed groups (p<0.05). Conclusion: We conclude that the balanced release of AA from CAS containing diets and mixed diets could reduce muscle degradation, favor nitrogen retention, % intake and improve growth performance in pigs consuming either a normal- or low-protein diet.

임신일령에 따른 생쥐 태아 뇌조직의 단백질 발현 양상 분석 (Analysis of brain protein expression in developing mouse fetus)

  • 한영훈;김홍래;조운비;우제석;진동일
    • 농업과학연구
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    • 제38권1호
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    • pp.65-70
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    • 2011
  • Development of mouse fetus brains can be defined morphologically and functionally by three developmental stages, embryo day (ED) 16, postnatal stage one week and eight weeks. These defined stages of brain development may be closely associated with differential gene expression rates due to limited cellular resources such as energy, space, and free water. Complex patterns of expressed genes and proteins during brain development suggests the changes in relative concentrations of proteins rather than the increase in numbers of new gene products. This study was designed to evaluate early protein expression pattern in mouse fetus brain. The mouse brain proteome of fetus at ED 15.5, and 19.5 was obtained using 2-dimensional gel electrophoresis (DE). Analysis of the 2-DE gels in pH 3-10 range revealed the presence of 15 differentially expressed spots, of which 11 spots were identified to be known proteins following MALDI-TOF analysis; 3 spots were up-regulated and 8 spots were down-regulated in the mouse fetus brain at ED 15.5. UP-regulated proteins were identified as MCG18238, isoform M2 of pyruvate kinase isozymes M1/M2, isoform 2 of heterogeneous nuclear ribonucleoprotein K, heterogeneous nuclear ribonucleoprotein H2, creatine kinase B-type, 40S ribosomal protein SA and hemoglobin subunit beta-H1. Down-regulated proteins were putative uncharacterized protein, lactoylglutathione lyase and secreted acidic cysteine rich glycoprotein. Our results revealed composite profiles of mouse fetus brain proteins related to mouse fetus development by 2-DE analysis implying possible roles of these proteins in neural differentiation.