• Title/Summary/Keyword: NS5A

Search Result 713, Processing Time 0.025 seconds

Selection and Target-Site Mapping of Peptides Inhibiting HCV NS5B Polymerase Using Phage Display

  • Kim, Min-Soo;Park, Chan-Hee;Lee, Jong-Ho;Myung, Hee-Joon
    • Journal of Microbiology and Biotechnology
    • /
    • v.18 no.2
    • /
    • pp.328-333
    • /
    • 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.

Thermal Shock Resistance of Bilayered YSZ Thermal Barrier Coating

  • Lee, Dong Heon;Kim, Tae Woo;Lee, Kee Sung;Kim, Chul
    • Journal of the Korean Ceramic Society
    • /
    • v.55 no.5
    • /
    • pp.452-460
    • /
    • 2018
  • This study investigate changes in mechanical behaviors such as indentation load-displacement and hardness of thermal barrier coatings (TBCs) using cycling of thermal shock test. Relatively dense and porous TBCs on nickel-based bondcoat/super alloy are prepared using different starting granules, 204C-NS and 204NS commercial powers, and the effect of double layers of 204C-NS on 204NS and 204NS on 204C-NS are investigated. The highest temperature applied during thermal shock test is $1100^{\circ}C$ and the maximum number of cycles is 1,200. The results indicate that bilayered TBC showed a relatively mechanically resistant property during thermal shock cycles and that the mechanical behavior is influenced by the microstructure of TBCs by exposure to high temperature during tests or different starting granules.

A Study on the Radio Wave Propagation Model in IMT-2000 (IMT-2000 주파수대역에서의 전파-전파 모델에 관한 연구)

  • Ra, Yoo-Chan;Lee, Seung-Woo;Shin, Hong-Kyu
    • The Journal of Korean Institute of Communications and Information Sciences
    • /
    • v.28 no.4A
    • /
    • pp.224-231
    • /
    • 2003
  • In this in this thesis, we have proposed the Kor-231 which compared with Hata by dividing 4 classes and 8 details, which based on that the area subdivision which is defined by the City planning Regulation of Korea classified on its rate of building-to-land, floor area and distance between buliding and botanical coefficient. The experiments are carried out at the point of the RMS delay spread and the recerived power in the two kinds of geographical areas, LOS(Line of Sight) and N-LOS(Non Line of Sight). When the measured result is compared with Kor-231 model, we can catch the result that received power are 25.5dB and 14.5dB, the RMS delay spread are 101ns and 35ns and N-LOS received power are 4.1dB and 1.6dB. So we have certified that it is well due to the result analyzed into the difference of the RMS delay spread from 74ns to 200ns.

Zika Virus Proteins NS2A and NS4A Are Major Antagonists that Reduce IFN-β Promoter Activity Induced by the MDA5/RIG-I Signaling Pathway

  • Ngan, Nguyen Thi Thuy;Kim, Seong-Jun;Lee, Jeong Yoon;Myoung, Jinjong
    • Journal of Microbiology and Biotechnology
    • /
    • v.29 no.10
    • /
    • pp.1665-1674
    • /
    • 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.

Dengue Virus 2 NS2B Targets MAVS and IKKε to Evade the Antiviral Innate Immune Response

  • Ying Nie;Dongqing Deng;Lumin Mou;Qizhou Long;Jinzhi Chen;Jiahong Wu
    • Journal of Microbiology and Biotechnology
    • /
    • v.33 no.5
    • /
    • pp.600-606
    • /
    • 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.

Interaction of Stomatin with Hepatitis C Virus RNA Polymerase Stabilizes the Viral RNA Replicase Complexes on Detergent-Resistant Membranes

  • Kim, Jung-Hee;Rhee, Jin-Kyu;Ahn, Dae-Gyun;Kim, Kwang Pyo;Oh, Jong-Won
    • Journal of Microbiology and Biotechnology
    • /
    • v.24 no.12
    • /
    • pp.1744-1754
    • /
    • 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.

Analysis of SCR, MVSCR, LVTSCR With I-V Characteristic and Turn-On-Time (SCR, MVSCR, LVTSCR의 Turn-on time 및 전기적 특성에 관한 연구)

  • Lee, Joo-Young
    • Journal of IKEEE
    • /
    • v.20 no.3
    • /
    • pp.295-298
    • /
    • 2016
  • In this paper, we analysed the properties of the conventional ESD protection devices such as SCR, MVSCR, LVTSCR. The electrical characteristics and the turn-on time properties are simulated by Synopsys T-CAD simulator. As the results, the devices have the holding voltages between 2V and 3V, and the trigger voltage of about 20V with SCR, of about 12V with MVSCR, of about 9V with LVTSCR. The results of the simulation for the turn-on time properties are 2.8ns of SCR, 2.2ns of MVSCR, 2.0ns of LVTSCR. Thus, we prove that LVTSCR has the shortest turn-on time. However, the second breakdown currents(It2) of the devices are 7.7A of SCR, 5.5A of MVSCR, 4A of LVTSCR. This different properties have to be adapted by the operation voltages for I/O Clamps.

In vitro Study of Nucleostemin as a Potential Therapeutic Target in Human Breast Carcinoma SKBR-3 Cells

  • Guo, Yu;Liao, Ya-Ping;Zhang, Ding;Xu, Li-Sha;Li, Na;Guan, Wei-Jun;Liu, Chang-Qing
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.5
    • /
    • pp.2291-2295
    • /
    • 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.

The Fluorescence Behavior of the Responsive Macrocycle by Aromatic Imine Molecules

  • Choi, Chang-Shik;Jeon, Ki-Seok;Lee, Ki-Hwan
    • Journal of Photoscience
    • /
    • v.11 no.32
    • /
    • pp.71-74
    • /
    • 2004
  • The macrocycle L exhibited a switch on-off behavior through the fluorescent responses by aromatic imine molecule 1 (X=H) / trifluoroacetic acid (TFA). In the 'switch on' state, it was supposed that the aromatic imine molecule 1 is in the cavity of macrocycle L and a photoinduced electron transfer (PET) from the nitrogen of azacrown part to the anthryl group is inhibited by the interaction between the aromatic imine molecule 1 and the azacrown part of macrocycle L. In the 'switch off' state, it was supposed that the protonated imine molecule 1 is induced by the continuous addition of TFA and a repulsion between the protonated azacrown part and the protonated imine molecule 1 is occurred. It was considered that this process induces the intermolecular PET from the protonated imine molecule 1 to the anthryl group of macrocycle L because of a proximity effect between the anthryl group and the protonated imine molecule 1. From the investigation of the transient emission decay curve, the macrocycle L showed three components (3.45 ns (79.72%), 0.61 ns (14.53%), and 0.10 ns (5.75%). When the imine molecule 1 was added in the macrocycle L as molar ratio=1:1, the first main component showed a little longer lifetime as 3.68 ns (82.75%) although the other two components were similar as 0.64 ns (14.28%) and 0.08 ns (2.96%). On the contrary, when the imine molecule 3 (X=C1) was added in the macrocycle L as molar ratio=l:1, all the three components were decreased such as 3.27 ns (69.83%), 0.44 ns (13.24%), and 0.06 ns (16.93%). The fluorescent pH titration of macrocycle L was carried out from pH=3 to pH=9. The macrocycle L and C $U^{2+}$- macrocycle L complex were intersected at about pH=5, while the E $u^{3+}$ -macrocycle L complex was intersected at about pH=5.5. In addtion, we investigated the fluorescence change of macrocycle L as a function of the substituent constant ($\sigma$$_{p}$$^{o}$) showing in the para-substituent with electron withdrawing groups (X=F, Cl) and electron donating groups (X=C $H_3$, OC $H_3$, N(C $H_3$)$_2$), respectively, as well as non-substituent (X=H).).ctively, as well as non-substituent (X=H).

  • PDF

40-W 200-ns 300-kHz Thulium-doped Fiber Laser at 2050 nm

  • Shin, Jae Sung;Cha, Yong-Ho;Chun, Byung Jae;Park, Hyunmin
    • Current Optics and Photonics
    • /
    • v.5 no.5
    • /
    • pp.544-553
    • /
    • 2021
  • A 40-W 200-ns 300-kHz thulium-doped fiber laser at 2050 nm with a master oscillator power amplifier configuration was developed, for application to lithium-isotope separation. The master oscillator generated a 5.35 W continuous-wave beam, which the pulse generator then broke into 200-ns pulses at 300 kHz. Then, the laser beam was amplified by passing through a two-stage amplifier. The output power finally obtained was 42.0 W at 2050 nm, and was stable for a long time, over 2 hours. In spite of this achievement, mode instability was observed in the output beam. This can be solved in the future by using a method such as tight coiling.