국내 분리 일본뇌염 바이러스 유전자에 대한 항체생성 연구(Ⅱ) : 일본뇌염 바이러스 유전자 발현 expression of JEV viral genes

주영란,최택균,유정희,이한샘,남재환,최우영,박근용,조해월 국립보건연구원 2000 국립보건원보 Vol.37 No.-

Interaction Study of Soybean mosaic virus Proteins with Soybean Proteins usingthe Yeast-Two Hybrid System

서장균,황성현,강성환,최홍수,이수헌,손승한,김국형 한국식물병리학회 2007 Plant Pathology Journal Vol.23 No.4

Interactions between viral proteins and host proteins are essential for virus replication. Especially, translation of viral genes completely depends on the host machinery. In potyviruses, interactions of genome-linked viral protein (VPg) with host translation factors including eIF4E, eIF(iso)4E, and poly(A)-binding protein (PABP) has previously been characterized. In this study, we investigated interactions between Soybean mosaic virus (SMV) viral proteins and host translation factors by yeast two-hybrid system. SMV VPg interacted with eIF4E, eIF(iso)4E, and PABP in yeast two-hybrid system, while SMV helper component proteinase (HCpro) interacted with neither of those proteins. The interaction between SMV NIb and PABP was also detected. These results are consistent with those reported previously in other potyviruses. Interestingly, we found reproducible and specific interactions between SMV coat protein (CP) and PABP. Deletion analysis showed that the region of CP comprising amino acids 116 to 206 and the region of PABP comprising amino acids 520 to 580 are involved in CP/PABP interactions. Soybean library screening with SMV NIb by yeast twohybrid assay also identified several soybean proteins including chlorophyll a/b binding preprotein, photosystem I-N subunit, ribulose 1,5-biphosphate carboxylase, ST-LS1 protein, translation initiation factor 1, TIRNBS type R protein, RNA binding protein, ubiquitin, and LRR protein kinase. Altogether, these results suggest that potyviral replicase may comprise a multiprotein complex with PABP, CP, and other host factors.

Kaposi’s Sarcoma-Associated Herpesvirus Viral Protein Kinase Interacts with RNA Helicase A and Regulates Host Gene Expression

Jae Eun Jong,박준수,Sunmi Kim,서태근 한국미생물학회 2010 The journal of microbiology Vol.48 No.2

RNA helicase A (RHA) containing the DExH motif is a human homolog of maleless protein that regulates expression of genes located in the Drosophila X chromosome during dosage compensation. RHA exerts helicase activity that unwinds double-stranded RNA and DNA to a single-strand form. The protein acts as a bridging factor mediating interactions of CBP/p300 and RNA pol II, and consequently affects gene expression. Kaposi’s sarcoma-associated herpesvirus (KSHV) is a member of the γ-herpesvirus subfamily that causes several disorders. The majority of herpesviruses commonly encode predicted viral protein kinases. KSHV open reading frame 36 (ORF36) codes for protein kinase domains, and functions as a serine/threonine protein kinase. KSHV ORF36 is classified as a late gene, as it is expressed during lytic replication and localized in the nuclei of KSHV-infected cells. Recent studies show that viral protein kinase (vPK) interacts with cellular proteins. In this study, we determined the cellular localization of vPK in KSHVinfected BCBL-1 cells using confocal microscopy. Proteomic analysis indicates that cellular proteins interacted with vPK, and co-immunoprecipitation reactions further reveal interactions between vPK and RHA. Moreover, KSHV vPK appeared to regulate the transcriptional activation of Cre promoter, and plays an important role in cellular transcription of RHA.

Characterization of the Putative Membrane Fusion Peptides in the Envelope Proteins of Human Hepatitis B Virus

Kang, Ha-Tan,Yu, Yeon-Gyu Korean Chemical Society 2007 Bulletin of the Korean Chemical Society Vol.28 No.10

Envelope proteins of virus contain a segment of hydrophobic amino acids, called as fusion peptide, which triggers membrane fusion by insertion into membrane and perturbation of lipid bilayer structure. Potential fusion peptide sequences have been identified in the middle of L or M proteins or at the N-terminus of S protein in the envelope of human hepatitis B virus (HBV). Two 16-mer peptides representing the N-terminal fusion peptide of the S protein and the internal fusion peptide in L protein were synthesized, and their membrane disrupting activities were characterized. The internal fusion peptide in L protein showed higher activity of liposome leakage and hemolysis of human red blood cells than the N-terminal fusion peptide of S protein. Also, the membrane disrupting activity of the extracellular domain of L protein significantly increased when the internal fusion peptide region was exposed to N-terminus by the treatment of V8 protease. These results indicate that the internal fusion peptide region of L protein could activate membrane fusion when it is exposed by proteolysis.

Characterization of the Putative Membrane Fusion Peptides in the Envelope Proteins of Human Hepatitis B Virus

Ha-Tan Kang,유연규 대한화학회 2007 Bulletin of the Korean Chemical Society Vol.28 No.10

Envelope proteins of virus contain a segment of hydrophobic amino acids, called as fusion peptide, which triggers membrane fusion by insertion into membrane and perturbation of lipid bilayer structure. Potential fusion peptide sequences have been identified in the middle of L or M proteins or at the N-terminus of S protein in the envelope of human hepatitis B virus (HBV). Two 16-mer peptides representing the N-terminal fusion peptide of the S protein and the internal fusion peptide in L protein were synthesized, and their membrane disrupting activities were characterized. The internal fusion peptide in L protein showed higher activity of liposome leakage and hemolysis of human red blood cells than the N-terminal fusion peptide of S protein. Also, the membrane disrupting activity of the extracellular domain of L protein significantly increased when the internal fusion peptide region was exposed to N-terminus by the treatment of V8 protease. These results indicate that the internal fusion peptide region of L protein could activate membrane fusion when it is exposed by proteolysis.

Viral Interferon Regulatory Factor 1 of Kaposi’s Sarcoma-Associated Herpesvirus Interacts with a Translocation Liposarcoma Protein-Associated Serine-Arginine Protein

Kim, Sunmi,Jong, Jae Eun,Seo, Taegun Elsevier 2012 Osong Public Health and Research Persptectives Vol.3 No.1

<P><B>Abstract</B></P><P><B>Objectives</B></P><P>To confirm that Kaposi’s sarcoma-associated herpes virus open-reading frame K9, viral interferon regulatory factor 1 (vIRF1), interacts with splicing factor, translocation liposarcoma protein-associated serine-arginine protein (TASR), <I>in vivo</I> and to establish whether interactions between vIRF1 and TASRs influence alternative splicing.</P><P><B>Methods</B></P><P>Association between vIRF1 and TASRs was confirmed with the glutathione S-transferase pull-down assay and coimmunoprecipitation. Further colocalization was shown by immunofluorescence. The <I>in vivo</I> splicing assay was performed to confirm the alterations in the splicing pattern.</P><P><B>Results</B></P><P>vIRF1 interacts with both TASR1 and 2 <I>in vivo</I>. vIRF1 has been shown to colocalize with TASR proteins in 293 T cells. However, an <I>in vivo</I> splicing revealed no alterations in the splicing pattern via interaction.</P><P><B>Conclusions</B></P><P>The study data suggest that vIRF1 interacts with the TASR protein. However, vIRF1 interactions do not affect TASR-mediated alternative splicing.</P>

소 설사병 바이러스 구조단백질 E2의 대장균 발현

권창희,윤용덕,안수환 대한바이러스학회 1994 Journal of Bacteriology and Virology Vol.24 No.1

The gene corresponding envelope(E2, C-terminal of El and E2) of bovine viral diarrhea virus (BVDV) was expressed in downstream from the malE gene of E. coli, resulting in the expression of fusion protein with maltose-binding protein(MBP). The expressed MBP-E2 fusion protein was affinity purified for the production of antiserum and the expression of E2 was identified by immunoblotting with antiserum against MBP E2 fusion protein. Although the protein band with molecular weight above 88 kDa was identified, the cell expressing C-terminal of El and E2 showed the degradation of expressed fusion protein. However, low molecular weight protein bands including MBP were frequently detected in immunoblotting assay, thus, indicating that C-terminal of El can play a role of signal peptide after translation in similar pattern of eukaryotic cell.

Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation

Choi, Jae-Woong,Kim, Jong-Wook,Nguyen, Lap P.,Nguyen, Huu C.,Park, Eun-Mee,Choi, Dong Hwa,Han, Kang Min,Kang, Sang Min,Tark, Dongseob,Lim, Yun-Sook,Hwang, Soon B. Korean Society for Molecular and Cellular Biology 2020 Molecules and cells Vol.43 No.5

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.

Further characterization of the causative virus of rabbit viral hepatitis, so-called rabbit haemorrhagic disease in Korea

정종식,정규식,이차수,신태균,Jyeong, Jong-sik,Jeong, Kyu-sik,Lee, Cha-soo,Shin, Tae-kyun The Korean Society of Veterinary Science 1992 大韓獸醫學會誌 Vol.32 No.3

국내에서 발생한 토끼 바이러스성 간염 소위 토끼 출혈병의 원인 바이러스를 감염토끼의 간조직으로 부터 분리정제한 후 바이러스의 핵산과 구성 단백질의 특징을 관찰하였던 바 다음과 같은 결과를 얻었다. 토끼간염바이러스는 분자량이 약 54 kilodalton인 한개의 구조단백을 가진 RNA 바이러스이며 바이러스 핵산의 크기는 약 7.5 kilobases로 나타났고 바이러스의 RNA는 배양세포에서는 감염을 일으키지 않았다. 바이러스 구성단백의 양상과 핵산의 크기 등을 종합해 볼 때 토끼의 간염 바이러스는 Caliciviridae에 속하는 것으로 간주된다. The causative virus causing rabbit hepatitis has been further characterized by evaluating viral proteins and viral nucleic acids of purified viruses from the liver of the experimentally infected rabbits. Rabbit hepatitis virus has one major structural protein of 54 kilodaltons and some minor proteins. Vrial RNA was resistant to DNAse I. The size of viral nucleic acid of this virus was calculated to be about 7.5 kilobases. These findings indicate that rabbit hepatitis virus belongs to the family Caliciviridae.

Nucleocapsid and Spike Proteins of SARS-CoV-2 Drive Neutrophil Extracellular Trap Formation

윤영진,이유빈,김선화,진희경,배재성,홍창원 대한면역학회 2021 Immune Network Vol.21 No.2

Patients with severe coronavirus disease 2019 (COVID-19) demonstrate dysregulated immune responses including exacerbated neutrophil functions. Massive neutrophil infiltrations accompanying neutrophil extracellular trap (NET) formations are also observed in patients with severe COVID-19. However, the mechanism underlying severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-induced NET formation has not yet been elucidated. Here we show that 2 viral proteins encoded by SARS-CoV-2, the nucleocapsid protein and the whole spike protein, induce NET formation from neutrophils. NET formation was ROS-independent and was completely inhibited by the spleen tyrosine kinase inhibition. The inhibition of p38 MAPK, protein kinase C, and JNK signaling pathways also inhibited viral protein-induced NET formation. Our findings demonstrate one method by which SARS-CoV-2 evades innate immunity and provide a potential target for therapeutics to treat patients with severe COVID-19.