• Title/Summary/Keyword: Ribonucleoprotein

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Lipoteichoic Acid Isolated from Lactobacillus plantarum Inhibits Melanogenesis in B16F10 Mouse Melanoma Cells

  • Kim, Hye Rim;Kim, Hangeun;Jung, Bong Jun;You, Ga Eun;Jang, Soojin;Chung, Dae Kyun
    • Molecules and Cells
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    • v.38 no.2
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    • pp.163-170
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    • 2015
  • Lipoteichoic acid (LTA) is a major component of the cell wall of Gram-positive bacteria. Its effects on living organisms are different from those of lipopolysaccharide (LPS) found in Gram-negative bacteria. LTA contributes to immune regulatory effects including anti-aging. In this study, we showed that LTA isolated from Lactobacillus plantarum (pLTA) inhibited melanogenesis in B16F10 mouse melanoma cells. pLTA reduced the cellular activity of tyrosinase and the expression of tyrosinase family members in a dose-dependent manner. The expression of microphthalmia- associated transcription factor (MITF), a key factor in the synthesis of melanin, was also decreased by pLTA. Further, we showed that pLTA activated melanogenesis signaling, such as extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3-kinse (PI3K)/AKT. In addition, the expression of heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) and HuR, which are important RNA-binding proteins (RBPs), was reduced. pLTA likely degrades MITF via regulation of melanogenic signaling and RNA stability of melanogenic proteins, resulting in the reduction of melanin. Thus, our data suggest that pLTA has therapeutic potential for treating hyperpigmentation disorders and can also be used as a cosmetic whitening agent.

Cloning of hnRNP E1 cDNA via yeast two-hybrid system and a study on protein-protein interaction between hnRNP E1 and hnRNP K (이스트 two-hybrid 시스템을 이용한 hnRNP E1 cDNA의 클로닝과 hnRNP E1-hnRNP K 상호결합에 대한 연구)

  • Choi, Mie-Young
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.9 no.6
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    • pp.1795-1799
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    • 2008
  • The heterogeneous nuclear ribonucleoprotein K (hnRNP K) is a component of hnRNP complexes. This protein binds strongly to cytidine-rich RNA/DNA sequences. It is a nucleocytoplasmic shuttling protein. To investigate the functions of hnRNP K, I searched for hnRNP K-interacting proteins in HeLa cDNA library using a yeast two-hybrid screening system. One of the cDNA clones is identical to human hnRNP E1 (poly(rC) binding protein 1) cDNA (GenBank accession number XM_031585). In this study, hnRNP K is shown to specifically interact with hnRNP E1 in yeast two-hybrid system and in vitro biochemical assay.

Expression of Telomerase Activity and Apoptosis in Human Brain Tumors (인체 뇌종양조직에서 텔로머레이즈의 발현과 세포사멸)

  • Kim, Choong Hyun;Cheong, Jin Hwan;Bak, Koang Hum;Kim, Jae Min;Ko, Yong;Oh, Suck Jun
    • Journal of Korean Neurosurgical Society
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    • v.30 no.2
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    • pp.137-143
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    • 2001
  • Objective : Telomerase, a ribonucleoprotein adds telomere repeats to the ends of telomeres to compensate for the progressive loss. A favorable prognosis associated with low or no telomerase activity in some tumors, and cells transfected with antisense human telomerase lost telomeric repeats and die. We studied about the relationship between telomerase activity and apoptosis in the human brain tumors. Material and Methods : Between July 1998 and December 1999, 62 patients with brain tumors underwent surgery and their surgical specimens were obtained. Telomerase activity was investigated by telomeric repeats amplification protocol(TRAP) assay. Apoptosis was also evaluated by DNA fragmentation analysis. Differences and correlation in data were analyzed using Mann-Whitney test and Wilcoxon-signed rank test. Results : Expression rate of telomerase activity and apoptosis were 80% and 30% in malignant gliomas, 33% and 0% in low grade gliomas, 63% and 38% in meningiomas, 67% and 33% in pituitary adenomas, 33% and 33% in metastatic tumors, 67% and 17% in acoustic neurinomas, 100% and 100% in pineoblastomas, 100% and 0% in the hemangioblastoma, respectively. There was no significant difference of telomerase activity and apoptosis between histological types. But a significant difference was noted in the expression of telomerase activity between malignant gliomas and low grade gliomas(p = 0.022). Brain tumors with telomerase activity expressed the lower rate of apoptosis. A significant correlation was also found between telomerase activity and absence of apoptosis in the human brain tumors(p = 0.005). Conclusions : Our data suggests that telomerase may protect from apoptosis of the human brain tumors and also may play an important role in the biological malignancy of the gliomas.

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M6A reader hnRNPA2/B1 is essential for porcine embryo development via gene expression regulation

  • Kwon, Jeongwoo;Jo, Yu-Jin;Yoon, Seung-Bin;You, Hyeong-ju;Youn, Changsic;Kim, Yejin;Lee, Jiin;Kim, Nam-Hyung;Kim, Ji-Su
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.2
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    • pp.121-129
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    • 2022
  • Heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2/B1) is an N6-methyladenosine (m6A) RNA modification regulator and a key determinant of prem-RNA processing, mRNA metabolism and transportation in cells. Currently, m6A reader proteins such as hnRNPA2/B1 and YTHDF2 has functional roles in mice embryo. However, the role of hnRNPA2/B1 in porcine embryogenic development are unclear. Here, we investigated the developmental competence and mRNA expression levels in porcine parthenogenetic embryos after hnRNPA2/B1 knock-down. HhnRNPA2/B1 was localized in the nucleus during subsequent embryonic development since zygote stage. After hnRNPA2/B1 knock-down using double stranded RNA injection, blastocyst formation rate decreased than that in the control group. Moreover, hnRNPA2/B1 knock-down embryos show developmental delay after compaction. In blastocyste stage, total cell number was decreased. Interestingly, gene expression patterns revealed that transcription of Pou5f1, Sox2, TRFP2C, Cdx2 and PARD6B decreased without changing the junction protein, ZO1, OCLN, and CDH1. Thus, hnRNPA2/B1 is necessary for porcine early embryo development by regulating gene expression through epigenetic RNA modification.

The Adenylyl Cyclase Activator Forskolin Increases Influenza Virus Propagation in MDCK Cells by Regulating ERK1/2 Activity

  • Sang-Yeon Lee;Jisun Lee;Hye-Lim Park;Yong-Wook Park;Hun Kim;Jae-Hwan Nam
    • Journal of Microbiology and Biotechnology
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    • v.33 no.12
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    • pp.1576-1586
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    • 2023
  • Vaccination is the most effective method for preventing the spread of the influenza virus. Cell-based influenza vaccines have been developed to overcome the disadvantages of egg-based vaccines and their production efficiency has been previously discussed. In this study, we investigated whether treatment with forskolin (FSK), an adenylyl cyclase activator, affected the output of a cell-based influenza vaccine. We found that FSK increased the propagation of three influenza virus subtypes (A/H1N1/California/4/09, A/H3N2/Mississippi/1/85, and B/Shandong/7/97) in Madin-Darby canine kidney (MDCK) cells. Interestingly, FSK suppressed the growth of MDCK cells. This effect could be a result of protein kinase A (PKA)-Src axis activation, which downregulates extracellular signal-regulated kinase (ERK)1/2 activity and delays cell cycle progression from G1 to S. This delay in cell growth might benefit the binding and entry of the influenza virus in the early stages of viral replication. In contrast, FSK dramatically upregulated ERK1/2 activity via the cAMP-PKA-Raf-1 axis at a late stage of viral replication. Thus, increased ERK1/2 activity might contribute to increased viral ribonucleoprotein export and influenza virus propagation. The increase in viral titer induced by FSK could be explained by the action of cAMP in assisting the entry and binding of the influenza virus. Therefore, FSK addition to cell culture systems could help increase the production efficiency of cell-based vaccines against the influenza virus.

Analysis of the Amount of Telomeric DNA and Telomerase Activity on Preimplantation Mouse Embryoic Cells (마우스 수정란의 초기 배 발생단계별 Telomeric DNA의 양적 분석과 Telomerase 활성도 분석)

  • Kang M. Y.;Han M. S.;Lee S. C.;Kim J. H.;Sohn S. H.
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.1-7
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    • 2005
  • Telomeres consisting of (TTAGGG)n tandem repeat DNA sequences and associated proteins are essential for chromosome stability and related with cell senescence, apoptosis and cancer. The telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. This study was carried out to identify the distribution of telomeres on mouse chromosomes and also to analyze the amount of telomeres and telomerase activity of mouse embryos at early embryonic stages. Germ cells and early embryos from 1 cell to blastocyst stage were analyzed. The amount of telomeres was analyzed by quantitative fluorescence in situ hybridization technique(Q-FISH) using a human telomeric DNA probe, and telomerase activity was measured by telomeric repeat amplification protocol assay(TRAP). In results, the telomeres on mouse chromosomes were distributed at the ends of all autosomes and sex chromosomes. Although the quantity of telomeres varied among chromosomes, most of chromosomes had higher amount in q-arm telomeres than in p-arm telomeres. The results of Q-FISH indicated that the relative amount of telomeres of mouse embryos in each embryonic stage was approximately the same except the higher amount in blastocysts. Using TRAP assay on mouse embryos, telomerase activity was detected in all preimplantation stages from mature oocytes to blastocysts. Especially the telomerase activity was significantly increased at the morula and blastocyst stage. In conclusion, there may be a close association between the amount of telomeres and telomerase activity in early embryonic stages, and analysis of telomere quantity and telomerase activity on early development will be helpful for the investigation of embryogenesis and embryonic cell differentiation in mice.

Molecular Cloning of Novel Genes Specifically Expressed in Snailfish, Liparis tanakae (꼼치, Liparis tanakae에서 특이하게 발현되는 새로운 유전인자의 검색)

  • 송인선;이석근;손진기
    • Development and Reproduction
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    • v.4 no.1
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    • pp.67-77
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    • 2000
  • Snailfish usually lives at the bottom of the sea and showed typical retrogressive change with specialized tissue structures of skin and skeletons. In order to obtain the specific genes of snailfish, highly expressed in the body, we made subtracted cDNA library and analyzed 200 clones. Totally 200 clones were obtained and sequenced, and among them 62 clones were turned out to be homologous to the known gene, i.e., thioesterase (9), myosin (8), creatine kinase (7), skeletal alpha-actin (6), parvalbumin b (5), ribosomal protein (5), type I collagen (3), muscle troponin (3), dopamine receptor (2), histatin (2), and heat shock protein (2), cystatin (1), lectin (1), statherin (1), secretory carrier membrane protein (1), keratin type I (1), desmin (1), chloroplast (1), muscle tropomyosin (1), reticulum calcium ATPase (1), ribonucleoprotein (1). The remaining 138 clones were low homologous or non-redundant genes through Genbank search. Especially 5 clones were novel and specifically expressed in the body tissues of Snailfish by in situ hybridization. Therefore, we analysed these 5 clones to identify the C-terminal protein structures and motifs, and partly defined the roles of these proteins in comparison with the expression patterns by in situ hybridization. C9O-77, about 5000 bp, was supposed to be a matrix protein expressed strongly positive in epithelium, myxoid tissue, fibrous tissue and collagenous tissue. C9O-116, about 1500 bp, was supposed to be a transmembrane protein which was weakly expressed in the fibrous tissue, epithelium tissue, and myxoid tissue, but strong in muscle tissue. C9O-130, about 1200 bp, was supposed to be an intracytoplasmic molecule usually in the epithelial cells. C9O-161, about 2000 bp, was weakly expressed in epithelium, muscle tissue and myxoid tissue, but specially strong in epithelium. C9O-171, about 1000 bp, was supposed to be a transcription factor containing zinc finger like domain, which was intensely expressed in the epithelium, muscle tissue, fibrous tissue, and in collagenous tissue.

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Five Genes Regulated by Oryctes rhinoceros nudivirus Infection in the Intestinal Tube of Allomyrina dichotoma (장수풍뎅이(Allomyrina dichotoma)에 Oryctes rhinoceros nudivirus 감염에 의해서 유전자 발현이 조절되는 5개의 유전자)

  • Yoo, Bo-Kyung;Kwon, Kisang;Ko, Young Hwa;Lee, Eun Ryeong;Choi, Ji-Young;Kwon, O-Yu
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1336-1340
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    • 2016
  • Korean rhinoceros beetles (Allomyrina dichotoma), which can be found in broad-leaved forests in mountainous habitats and lives for around one year in wild. This beetle is currently popular as a pet and traditionally regarded as a medicine for liver-related diseases in Korea. It is reported that the economic losses in the mass-rearing facilities by virus infection have been increased since the 2010s in Korea. The causing virus for the A. dichotoma was firstly reported as an Oryctes rhinoceros nudivirus (OrNV) in 2015. We, here, observes that serious morphological changes in the intestinal tube from the OrNV-infected beetles, and report five genes, which are regulated by OrNV infection in the intestine; Krueppel-like factor 15 (Klf15), Endoplasmic reticulum aminopeptidase 2 (ERAP2), U5 small nuclear ribonucleoprotein 200 kDa helicase (Snrnp200), Muscleblind-like protein 2a (mbnl2a), and MIMI_L93. The results may provide a clue to the early diagnosis and disease treatment during the mass-rearing facilities of the A. dichotoma.

Ataxin-2 Dysregulation Triggers a Compensatory Fragile X Mental Retardation Protein Decrease in Drosophila C4da Neurons

  • Cha, In Jun;Lee, Davin;Park, Sung Soon;Chung, Chang Geon;Kim, Seung Yeon;Jo, Min Gu;Kim, Seung Yeol;Lee, Byung-Hoon;Lee, Young-Sam;Lee, Sung Bae
    • Molecules and Cells
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    • v.43 no.10
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    • pp.870-879
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    • 2020
  • Dendrites require precise and timely delivery of protein substrates to distal areas to ensure the correct morphology and function of neurons. Many of these protein substrates are supplied in the form of ribonucleoprotein (RNP) complex consisting of RNA-binding proteins (RBPs) and mRNAs, which are subsequently translated in distal dendritic areas. It remains elusive, however, whether key RBPs supply mRNA according to local demands individually or in a coordinated manner. In this study, we investigated how Drosophila sensory neurons respond to the dysregulation of a disease-associated RBP, Ataxin-2 (ATX2), which leads to dendritic defects. We found that ATX2 plays a crucial role in spacing dendritic branches for the optimal dendritic receptive fields in Drosophila class IV dendritic arborization (C4da) neurons, where both expression level and subcellular location of ATX2 contribute significantly to this effect. We showed that translational upregulation through the expression of eukaryotic translation initiation factor 4E (eIF4E) further enhanced the ATX2-induced dendritic phenotypes. Additionally, we found that the expression level of another disease-associated RBP, fragile X mental retardation protein (FMRP), decreased in both cell bodies and dendrites when neurons were faced with aberrant upregulation of ATX2. Finally, we revealed that the PAM2 motif of ATX2, which mediates its interaction with poly(A)-binding protein (PABP), is potentially necessary for the decrease of FMRP in certain neuronal stress conditions. Collectively, our data suggest that dysregulation of RBPs triggers a compensatory regulation of other functionally-overlapping RBPs to minimize RBP dysregulation-associated aberrations that hinder neuronal homeostasis in dendrites.

CLINICAL IMPLICATIONS OF TELOMERASE ACTIVITY IN ORAL SQUAMOUS CELL CARCIMOMA (구강편평세포암에서 telomerase 활성도의 임상적 연관성에 관한 연구)

  • Shim, Yu-Jin;Kim, Myung-Jin;Nahm, Dong-Seok;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.4
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    • pp.289-300
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    • 2001
  • Telomerase is a ribonucleoprotein that synthesizes telomere repeats. It has been reported that activation of telomerase was associtated with immortalization, proliferative activity and carcinogenesis. Recently, telomerase activity has been extensively studied in many kinds of malignant tumors for clinical diagnostic and/or prognostic utilities. In neuroblastoma, breast carcinoma, gastric carcinoma, non-small cell lung carcinoma, close relationship has been reported between high telomerase activity and lymph node metastasis, tumor aggressiveness and poor prognosis. The purpose of this study is to investigate the clinical implication of telomerase activity assay as an adjunctive factor in decision-making on neck node management, speedy pre-operative judging on histologic malignancy grading. Thus we performed semi-quantitative assay of telomerase activity using Telomerase PCR ELISA $kit^{(R)}$(Boeringer Manheim, Germany) and evaluated correlation between telomerase activity and tumor size, neck node metastasis, Anneroth malignancy score and influence of pre-operative chemotherapy on its activity in 27 cases of oral squamous cell carcinomas and 18 cases of normal oral epithelium. Also, correlation between telomerase activities and PCNA indices was evaluated. The results were obtained as follows: 1. The telomerase activities were detected in 24 specimens out of 27 oral squamous cell carcinoma specimens (88.9%) and in 5 specimens out of 18 normal oral epithelium specimens (27.8%). The mean value of telomerase activities was $0.9793{\pm}0.3428$ in 24 oral squamous cell carcinoma specimens and $0.4855{\pm}0.1117$ in 5 normal oral epithelium specimens. The positivity rate and mean value of telomerase activities in oral squamous cell carcinoma specimens were significantly higher than those of normal oral epithelium specimens (p<0.05). 2. There was no significant correlation between total Anneroth malignancy score and telomerase activity (p>0.05), but points of mitosis index and depth of invasion were significantly correlated with telomerase activities (p<0.05). 3. The positive immunohistochemical staining for PCNA(proliferating cell nuclear antigen) was observed in 26 specimens out of 27 oral squamous cell carcinoma specimens and mean value of PCNA indices of 26 specimens was $53.67{\pm}26.46$. PCNA indices were significantly correlated with telomerase activities (p<0.05). 4. The mean value of telomerase activities was significantly higher in pathologic T3/T4 group than in T1/T2 group (p<0.01). There was no significant difference of mean value of telomerase activities between pathologic neck node positive group and negative group (p> 0.05). Pre-operative chemotherapy significantly lowered the telomerase activities (p<0.05). The above results suggested telomerase activity could be used as diagnostic marker and adjunctive parameter for judging on histologic malignancy in oral squamous cell carcinoma.

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