• Molecules and Cells
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• 제43권5호
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• pp.469-478
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• 2020

Hepatitis C virus (HCV) propagation is highly dependent on cellular proteins. To identify the host factors involved in HCV propagation, we previously performed protein microarray assays and identified the LIM and SH3 domain protein 1 (LASP-1) as an HCV NS5A-interacting partner. LASP-1 plays an important role in the regulation of cell proliferation, migration, and protein-protein interactions. Alteration of LASP-1 expression has been implicated in hepatocellular carcinoma. However, the functional involvement of LASP-1 in HCV propagation and HCV-induced pathogenesis has not been elucidated. Here, we first verified the protein interaction of NS5A and LASP-1 by both in vitro pulldown and coimmunoprecipitation assays. We further showed that NS5A and LASP-1 were colocalized in the cytoplasm of HCV infected cells. NS5A interacted with LASP-1 through the proline motif in domain I of NS5A and the tryptophan residue in the SH3 domain of LASP-1. Knockdown of LASP1 increased HCV replication in both HCV-infected cells and HCV subgenomic replicon cells. LASP-1 negatively regulated viral propagation and thereby overexpression of LASP-1 decreased HCV replication. Moreover, HCV propagation was decreased by wild-type LASP-1 but not by an NS5A binding-defective mutant of LASP-1. We further demonstrated that LASP-1 was involved in the replication stage of the HCV life cycle. Importantly, LASP-1 expression levels were increased in persistently infected cells with HCV. These data suggest that HCV modulates LASP-1 via NS5A in order to regulate virion levels and maintain a persistent infection.

• 한국광학회지
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• 제14권5호
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• pp.515-520
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• 2003

수 KHz의 반복율로 발진하는 레이저광을 증폭하는 증폭기를 제작하고, 특성을 조사하였다. 증폭기로는 연속 방전하는 섬광등으로 펌핑되는 Nd:YAG 헤드를 사용하였고, 레이저 매질에서 발생하는 열 복굴절을 보상하여 이중통과 증폭기를 구성한 경우, 복굴절에 의한 레이저광 손실을 5%로 감소시켰다. 4 중통과 증폭기를 구성을 하였을 때, 반복율 5-10 KHz에서 이득은 약 3.2이었고, 펄스폭은 40 ns에서 48 ns로. 20%증가하였다.

• 대한전자공학회논문지TC
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• 제46권5호
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• pp.162-167
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• 2009

본 논문에서는 대역통과 여파기와 저역통과 여파기를 하나의 구조로 합성하여 크기 증가 없이 초광대역 특성을 가지는 여파기를 설계 및 구현하였다. 제안한 구조는 그 구조가 간단하다. 그리고 저역 통과 특성을 얻기 위하여 이미 잘 알려진 DGS(Defected Ground Structure) 구조를 이용하였다. 기존의 대역통과 여파기와 저역통과 여파기를 직접 연결하였을 때 보다 크기를 줄일 수 있었다. 측정결과 통과대역은 $2.1GHz{\sim}10.56GHz$이고 삽입손실은 최대 0.5dB, 반사손실은 최소 20dB, 군 지연은 $0.13ns{\sim}0.35ns$ 까지 최대 변화폭 0.23ns의 변화를 보인다.

• Journal of Microbiology and Biotechnology
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• 제18권2호
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• pp.328-333
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• 2008

A series of pep tides binding to the HCV NS5B polymerase was selected from phage display peptide libraries. A conserved motif of Ser-Arg-X-Arg/Leu was identified among the selected peptides, and Pep2 (Trp-Ser-Arg-Pro-Arg-Ser-Leu) was chosen for further characterization. The binding of Pep2 to HCV NS5B in vivo was shown by a yeast two-hybrid assay and by subcellular colocalization analysis using immunofluorescence confocal microscopy. The in vitro interaction was also confirmed by GST pulldown assay. The replication of the HCV 1b subgenomic replicon was efficiently inhibited by the presence of the peptide. By using a subtractive biopanning against Pep2, the binding site of the peptide was mapped at the pocket of Pro388 to Pro391 in the thumb subdomain of the polymerase. A yeast two-hybrid analysis using Pro388Ala and Pro391Ala mutants of NS5B confirmed the binding.

• 사상체질의학회지
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• 제16권3호
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• pp.96-107
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• 2004

1. Objectives This study was carried out to investigate the effects of Gunyuljejo-tang on the $CCl_4$-induced Liver Damage in Rats. 2. Methods Sprague-Dawley rats were devided into 5 experimental groups : Normal, $NS+CCl_4$(Solid extract of $CCl_4$ injection group after Normal Saline feed), $GYJJT+CCl_4$(Solid extract of $CCl_4$ injection group after Gunyuljejo-tang feed), $CCl_4+NS$(Normal Saline feed group after $CCl_4$ injection), $CCl_4+GYJJT$(Solid extract of Gunyuljejo-tang feed group after $CCl_4$ injection). Biochemical assays for serum enzyme activities such as AST, ALT, ALP, BUN, Creatinine, Uric Acid, Total Protein, Albumin, Total Cholesterol, Triglyceride, Glucose, and mRNA Revelation of Cytochrome p450 and activities such as LPO, GSH, GST, Glutathione Reductase, Glutathione Peroxidase, SOD, Catalase, Hydroxyproline, and ${\beta}$-Glucuronidase were performed. 3. Results (1) $GYJJT+CCl_4$ showed lower revelation of Cytochrome p450. (2) $GYJJT+CCl_4$ showed higher GSH activity than $NS+CCl_4$, $CCl_4+GYJJT$ showed higher GSH activity than $CCl_4+NS$ injection significantly. (3) $GYJJT+CCl_4$ showed higher GST activity than $NS+CCl_4$. $CCl_4+GYJJT$ showed higher GST activity than $CCl_4+NS$ significantly. (4) $GYJJT+CCl_4$ showed higher Glutathione Peroxidase activity than $NS+CCl_4$, $CCl_4+GYJJT$ showed higher Glutathione Peroxidase activity than $CCl_4+NS$ significantly. (5) $CCl_4+GYJJT$ showed higher SOD activity than $CCl_4+NS$ significantly. (6) $CCl_4+GYJJT$ showed higher Catalase activity than $CCl_4+NS$ significantly. (7) $GYJJT+CCl_4$ showed lower Hydroxyproline than $NS+CCl_4$ significantly, $CCl_4+GYJJT$ showed higher Hydroxyproline than $CCl_4+NS$ significantly. (8) $GYJJT+CCl_4$ showed higher ${\beta}$-Glucuronidase activity than $NS+CCl_4$, $CCl_4+GYJJT$ showed higher ${\beta}$-Glucuronidase activity than $CCl_4+NS$ significantly. 4. Conclusions Gunyuljejo-tang has the recovering effects on the $CCl_4$-induced Liver Damage significantly.

• Journal of Microbiology and Biotechnology
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• 제29권10호
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• pp.1665-1674
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• 2019

Zika virus (ZIKV) is a mosquito-transmitted, emerging Flavivirus that causes Guillain-$Barr{\acute{e}}$ syndrome and microcephaly in adults and fetuses, respectively. Since ZIKV was first isolated in 1947, severe outbreaks have occurred at various places worldwide, including Yap Island in 2007, French Polynesia in 2013, and Brazil in 2015. Although incidences of ZIKV infection and dissemination have drastically increased, the mechanisms underlying the pathogenesis of ZIKV have not been sufficiently studied. In addition, despite extensive research, the exact roles of individual ZIKV genes in the viral evasion of the host innate immune responses remain elusive. Besides, it is still possible that more than one ZIKV-encoded protein may negatively affect type I interferon (IFN) induction. Hence, in this study, we aimed to determine the modulations of the IFN promoter activity, induced by the MDA5/RIG-I signaling pathway, by over-expressing individual ZIKV genes. Our results show that two nonstructural proteins, NS2A and NS4A, significantly down-regulated the promoter activity of IFN-${\beta}$ by inhibiting multiple signaling molecules involved in the activation of IFN-${\beta}$. Interestingly, while NS2A suppressed both full-length and constitutively active RIG-I, NS4A had inhibitory activity only on full-length RIG-I. In addition, while NS2A inhibited all forms of IRF3 (full-length, regulatory domain-deficient, and constitutively active), NS4A could not inhibit constitutively active IRF3-5D. Taken together, our results showed that NS2A and NS4A play major roles as antagonists of MDA5/RIG-I-mediated IFN-${\beta}$ induction and more importantly, these two viral proteins seem to inhibit induction of the type I IFN responses in differential mechanisms. We believe this study expands our understanding regarding the mechanisms via which ZIKV controls the innate immune responses in cells and may pave the way to development of ZIKV-specific therapeutics.

• Asian Pacific Journal of Cancer Prevention
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• 제15권5호
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• pp.2291-2295
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• 2014

Although nucleolar protein nucleostemin (NS) is essential for cell proliferation and early embryogenesis and expression has been observed in some types of human cancer and stem cells, the molecular mechanisms involved in mediation of cell proliferation and cell cycling remains largely elusive. The aim of the present study was to evaluate NS as a potential target for gene therapy of human breast carcinoma by investigating NS gene expression and its effects on SKBR-3 cell proliferation and apoptosis. NS mRNA and protein were both found to be highly expressed in all detected cancer cell lines. The apoptotic rate of the pcDNA3.1-NS-Silencer group ($12.1-15.4{\pm}3.8%$) was significantly higher than those of pcDNA3.1-NS ($7.2-12.0{\pm}1.7%$) and non-transfection groups ($4.1-6.5{\pm}1.8%$, P<0.01). MTT assays showed the knockdown of NS expression reduced the proliferation rate of SKBR-3 cells significantly. Matrigel invasion and wound healing assays indicated that the number of invading cells was significantly decreased in the pcDNA3.1-NS-siRNA group (P<0.01), but there were no significant difference between non-transfected and over-expression groups (P>0.05). Moreover, RNAi-mediated NS down-regulation induced SKBR-3 cell G1 phase arrest, inhibited cell proliferation, and promoted p53 pathway-mediated cell apoptosis in SKBR-3 cells. NS might thus be an important regulator in the G2/M check point of cell cycle, blocking SKBR-3 cell progression through the G1/S phase. On the whole, these results suggest NS might be a tumor suppressor and important therapeutic target in human cancers.

• Journal of Microbiology and Biotechnology
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• 제33권5호
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• pp.600-606
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• 2023

Dengue virus (DENV) is a widespread arbovirus. To efficiently establish infection, DENV evolves multiple strategies to hijack the host innate immune response. Herein, we examined the inhibitory effects of DENV serotype 2 (DENV2) nonstructural proteins on RIG-I-directed antiviral immune response. We found that DENV2 NS2A, NS2B, NS4A, and NS4B significantly inhibited RIG-I-mediated IFN-β promoter activation. The roles of NS2B in RIG-I-directed antiviral immune response are unknown. Our study further showed that NS2B could dose-dependently suppress RIG-I/MAVS-induced activation of IFN-β promoter. Consistently, NS2B significantly decreased RIG-I- and MAVS-induced transcription of IFNB1, ISG15, and ISG56. Mechanistically, NS2B was found to interact with MAVS and IKKε to impair RIG-I-directed antiviral response. Our findings demonstrated a previously uncharacterized function of NS2B in RIG-I-mediated antiviral response, making it a promising drug target for anti-DENV treatments.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1744-1754
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• 2014

The hepatitis C virus (HCV) RNA genome is replicated by an RNA replicase complex (RC) consisting of cellular proteins and viral nonstructural (NS) proteins, including NS5B, an RNA-dependent RNA polymerase (RdRp) and key enzyme for viral RNA genome replication. The HCV RC is known to be associated with an intracellular membrane structure, but the cellular components of the RC and their roles in the formation of the HCV RC have not been well characterized. In this study, we took a proteomic approach to identify stomatin, a member of the integral proteins of lipid rafts, as a cellular protein interacting with HCV NS5B. Co-immunoprecipitation and co-localization studies confirmed the interaction between stomatin and NS5B. We demonstrated that the subcellular fraction containing viral NS proteins and stomatin displays RdRp activity. Membrane flotation assays with the HCV genome replication-competent subcellular fraction revealed that the HCV RdRp and stomatin are associated with the lipid raft-like domain of membranous structures. Stomatin silencing by RNA interference led to the release of NS5B from the detergent-resistant membrane, thereby inhibiting HCV replication in both HCV subgenomic replicon-harboring cells and HCV-infected cells. Our results identify stomatin as a cellular protein that plays a role in the formation of an enzymatically active HCV RC on a detergent-resistant membrane structure.

• 전기전자학회논문지
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• 제20권3호
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• pp.295-298
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• 2016

본 논문에서는 기존 ESD 보호소자인 SCR과 MVSCR, LVTSCR의 Turn-on-Time 및 전기적 특성을 시놉시스사의 T-Cad 시뮬레이션을 통하여 분석하였다. 분석결과 세 소자 모두 대략 2V 에서 3V 내외의 홀딩전압 특성을 보였으며, SCR은 약 20V의, MVSCR은 약 12V, LVTSCR은 9V로 순차적으로 개선된 트리거 특성을 보였다. 턴-온타임 시뮬레이션 결과는 SCR이 2.8ns, MVSCR과 LVTSCR은 각각 2.2ns, 2.0ns로 LVTSCR이 가장 짧은 턴-온 특성을 보였다. 반면 IT2 는 SCR이 약 7.7A, MVSCR은 5.5A LVTSCR은 4A의 특성을 보였으므로 I/O 및 파워 클램프 단에 적용 시 동작전압에 따른 최적화된 소자를 선택해야 한다.