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Molecular Miology of the Poliovirus (폴리오바이러스의 분자생물학)

  • 최원상
    • Journal of Life Science
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    • v.7 no.4
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    • pp.392-401
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    • 1997
  • The poliovirus is a small, and non-enveloped virus. The RNA genome of poliovirus is continuous, linear, and has a single open reading frame. This polyprotein precursor is cleaved proteolytically to yield mature products. Most of the cleavages occur by viral protease. The mature proteins derived from the P1 polyprotein precursor are the structural components of the viral capsid. The initial cleavage by 2A protease is indirectly involved in the cleavage of a cellular protein p220, a subunit of the eukaryotic translation initiation factor 4F. This cleavage leads to the shut-off of cap-dependent host cell translation, and allows poliovirus to utilize the host cell machinery exclusively for translation its own RNA, which is initiated by internal ribosome entry via a cap-independent mechanism. The functional role of the 2B, 2C and 2BC proteins are not much known. 2B, 2C, 2BC and 3CD proteins are involved in the replication complex of virus induced vesicles. All newly synthesized viral RNAs are linked with VPg. VPg is a 22 amino acid polypeptide which is derived from 3AB. The 3C and 3CD are protease and process most of the cleavage sites of the polyprotein precursor. The 3C protein is also involved in inhibition of RNA polymerase II and III mediated transcription by converting host transcription factor to an inactive form. The 3D is the RNA dependent RNA polymerase. It is known that poliovirus replication follows the general pattern of positive strand RNA virus. Plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA strands. Poliovirus RNA synthesis occurs in a membranous environment but how the template RNA and proteins required for RNA replication assemble in the membrane is not much known. The RNA requirements for the encapsidation of the poliovirus genome (packaging signal) are totally unknown. The poliovirus infection cycle lasts approximately 6 hours.

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Molecular Characterization and Expression Analysis of Clathrin-Associated Adaptor Protein 3-δ Subunit 2 (AP3S2) in Chicken

  • Oh, Jae-Don;Bigirwa, Godfrey;Lee, Seokhyun;Song, Ki-Duk
    • Korean Journal of Poultry Science
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    • v.46 no.1
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    • pp.31-37
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    • 2019
  • A chicken clathrin-associated adaptor protein $3-{\delta}$ subunit 2 (AP3S2) is a subunit of AP3, which is involved in cargo protein trafficking to target membrane with clathrin-coated vesicles. AP3S2 may play a role in virus entry into host cells through clathrin-dependent endocytosis. AP3S2 is also known to participate in metabolic disease developments of progressions, such as liver fibrosis with hepatitis C virus infection and type 2 diabetes mellitus. Chicken AP3S2 (chAP3S2) gene was originally identified as one of the differentially expressed genes (DEGs) in chicken kidney which was fed with different calcium doses. This study aims to characterize the molecular characteristics, gene expression patterns, and transcriptional regulation of chAP3S2 in response to the stimulation of Toll-like receptor 3 (TLR3) to understand the involvement of chAP3S2 in metabolic disease in chicken. As a result, the structure prediction of chAP3S2 gene revealed that the gene is highly conserved among AP3S2 orthologs from other species. Evolutionarily, it was suggested that chAP3S2 is relatively closely related to zebrafish, and fairly far from mammal AP3S2. The transcriptional profile revealed that chAP3S2 gene was highly expressed in chicken lung and spleen tissues, and under the stimulation of poly (I:C), the chAP3S2 expression was down-regulated in DF-1 cells (P<0.05). However, the presence of the transcriptional inhibitors, BAY 11-7085 (Bay) as an inhibitor for nuclear factor ${\kappa}B$ ($NF{\kappa}B$) or Tanshinone IIA (Tan-II) as an inhibitor for activated protein 1 (AP-1), did not affect the expressional level of chAP3S2, suggesting that these transcription factors might be dispensable for TLR3 mediated repression. These results suggest that chAP3S2 gene may play a significant role against viral infection and be involved in TLR3 signaling pathway. Further study about the transcriptional regulation of chAP3S2 in TLR3 pathways and the mechanism of chAP3S2 upon virus entry shall be needed.

Characterization of a cDNA Encoding Transmembrane Protein 258 from a Two-spotted Cricket Gryllus bimaculatus (쌍별귀뚜라미(Gryllus bimaculatus)의 GbTmem258 cDNA 클로닝과 발현분석)

  • Kisang Kwon;Honggeun Kim;Hyewon Park;O-Yu Kwon
    • Journal of Life Science
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    • v.33 no.10
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    • pp.828-834
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    • 2023
  • The cDNA that encodes transmembrane protein 258 (Tmem258) was cloned from Gryllus bimaculatus and named GbTmem258. This protein comprises 80 amino acids, has no N-glycosylation site, and contains five potential phosphorylation sites at two serines, two threonines, and one tyrosine. The predicted molecular mass of GbTmem258 is 9.06 kDa, and its theoretical isoelectric point is 5.5. The tertiary structure of GbTmem258 was predicted using the available secondary structure information, which suggests the presence of alpha helices (52.5%), random coils (22.5%), extended strands (16.25%), and beta turns (8.75%). Homology analysis revealed that GbTmem258 exhibits high similarity at the amino-acid level to Tmem258 found in other species. The effect of starvation and refeeding on GbTmem258 mRNA expression was also examined in this study. It was found that GbTmem258 mRNA expression in the hindgut progressively increased throughout the starvation period, peaking at almost 1.5 times the control level after six days of starvation. However, refeeding for one to two days after the six-day starvation period restored GbTmem258 mRNA expression to the control level. In fat body, GbTmem258 mRNA expression was almost 3-fold higher during starvation compared to the control level. Refeeding for one to two days after the six-day fast resulted in a decline in the expression to about a 2.5-fold increase over the control level. Throughout the starving and refeeding periods, no other tissues showed any discernible alterations in GbTmem258 mRNA expression.

Purification and Characterization of Lactate Dehydrogenase Isozymes in Channa argus (가물치(Channa argus) 젖산탈수소효소 동위효소들의 정제 및 특성)

  • Park, Eun-Mi;Yum, Jung-Joo
    • Journal of Life Science
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    • v.20 no.2
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    • pp.260-268
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    • 2010
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Channa argus were purified and characterized by biochemical, immunochemical and kinetic methods. The activity of LDH in skeletal muscle was the highest at 380.4 units and those in heart, eye and brain tissues were 13.4, 3,5 and 5.4 units, respectively. Citrate synthase (EC 4.1.3.7, CS) activity in heart tissue was the highest at 20.7 units. LDH/CS in skeletal muscle, heart, eye and brain tissues were 172.9, 0.6, 0.32 and 0.47. Protein concentration in skeletal muscle tissue was 14.7 mg/g and specific activities of LDH in skeletal muscle, heart, eye and brain tissues were 25.88, 0.79, 0.31 and 1.38 units/mg, respectively. Therefore, skeletal muscle tissue was anaerobic and heart tissue was aerobic. The LDH isozymes in tissues were identified by polyacrylamide gel electrophoresis, immunoprecipitation and Western blot with antiserum against $A_4$, $B_4$, and eye-specific $C_4$. LDH $A_4$, $A_3B$, $A_2B_2$. $AB_3$ and $B_4$ isozymes were detected in every tissue, $C_4$, $AC_3$, $A_2C_2$ and $A_3C$ were detected in eye tissue, and $A_3C$ was found in brain tissue. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ isozymes were purified by affinity chromatography and Preparative PAGE Cells. The LDH $A_4$ isozyme was purified in the fraction from elution with $NAD^+$ containing buffer of affinity chromatography. Eye-specific $C_4$ isozyme was eluted right after $A_4$, after which $B_4$ isozyme was eluted with plain buffer. As a result, one part of molecular structures in $A_4$, $B_4$ and eye-specific $C_4$ were similar, but were different from each other in $B_4$ and $C_4$. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The activity of LDH $A_4$, $A_2B_2$, $B_4$, and eye-specific $C_4$ isozymes remained at 39.98, 21.28, 19.67 and 16.87% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The $Km^{PYR}$ values were 0.17, 0.27 and 0.133 mM in $A_4$, $B_4$ and eye-specific $C_4$ isozymes, respectively. The optimum pH of LDH $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$, and $AB_3$ were pH 6.5, pH 8.5, pH 5.5, pH 6.0-6.5, pH 5.0 and pH 7.5. The $A_4$ and heterotetramer isozymes stabilized a broad range of pH. Especially, LDH activities in skeletal muscle tissue were high, resulting in a high degree of muscle activity.LDH metabolism in eye tissue seems to be converted faster from pyruvate to lactate by eye-specific $C_4$ isozyme as eye-specific $C_4$ have the highest affinity for pyruvate, and right after the conversion, oxidation of lactate was induced by $A_4$ isozyme. It was found that expression of Ldh-C, affinity to substrate and reaction time of $C_4$ isozyme were different according to the ecological environmental and feeding capturing patterns.

Ecological Studies on the Forest Vegetation in the Mt. Joghe (조계산(曹溪山) 삼림식생(森林植生)의 생태학적(生態學的) 연구(硏究))

  • Chang, Seok Mo
    • Journal of Korean Society of Forest Science
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    • v.80 no.1
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    • pp.54-71
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    • 1991
  • To classify and analyze the forest communities and their structures, the vegetation in Mt. Joghe was investigated from July, 1980 to August, 1989. The results obtained are as follow ; 1. A total of 750 kinds of vascular plant(49 orders, 122 families, 434 genera, 627 species, 1 subspecies, 111 varieties and 11 forma)were observed in Mt. Ioghe. The newly observed plant species were Dioscorea quingueloba, Spiranthes sinensis, Cephalanthera falcata, Angelica gigas, Clematis patents, Paeonia obovata, Hibiscus mulabilis, Ainsliaea acerifolia, Dictamnus dasycarpus, Cynachum ascyrifolia, Vaccinium koreanum, Erythrortium japonicum, Indigofera kirilowii (17species), Broussonetia kazinoki var, humillis, Euonymus, fortunei var. radicans, Juniperus communis var, nippnnica, Callicarpa japonica var. radicans, Joniperus communis var. rzipponica, Callicarpa japonica var. taquetii (4 varieties) and L indera obtusiloba for. billosum (1 forma). 2. The life spectrum of flora in Mt. Joghe was classified into $CH-D_1-R_5-e$ type. Distribution area was identical to Southern type by Nakai, Lee, and Yim. A few subtropical species were also observed. 3. Simpson's species diversity index(Ds) was 0.9 and Shannon-Weiner's diversity index (H') was 1.004. These indice suggest that the vegetation in Mt. Joghe is of complicated forest communities. 4. Pte-Q was 1.81 which was higher than the nationwide mean of 1.68. Urbanization Index (UI) was 28.75 for naturalized plant species, and 17.49 for exotic woody plant species, which were similar to those of Mt. Baekun and Mt. Naejang. 5. The forest vegetation of Mt. Joghe was grouped in 3 vegetation types : 7 natural plant Communities dominated by Quercus serrat, Quercus acutissima, Quercus variabilis, Carpinus laxiflora, Pinus derasiflora and Platycarya strobilacea, 8 substitutional plant communities Styrax japonica, Stewartia koreana, Lindera erytlrrocarpa, Zelkova serrata, Rhtrs chinensis, Controversa, and Frzrxirtus manrlshurica, and 7 plantation Communities composed of Pinus koraiensis, Pinus rigida, Magnolia nbnvata, Chamecyparis obkrsa, Larie ieptolepis, Castanea crenata and Cryptomeria japonica. 6. Actual vegetation maps and profile diagrams were made by phytosocialogical classification. 7. As the important and unique species in Mt. Joghe, Lindera sericea, Penicaria tilitorme, Hex macropoda, Hex macropoda for. pseudo-macropoda, Steroartia koreana, Adenopkora palustris and Corylop.,is coreana, which were also seported by Lee(1977), Kim and Yark(1989), were identified and Vaccinium coreanum, Cremastra appendiculinium, Juniperus comminis van. nipponica, Cephalanthera falcata, Broussortetia kazinoki var. humilis, paeonia obovata, Deutzia prunifolia, Dictamnus dasyarpus, Angelica gigics and Bupleurum falcatum were odditionally observed.

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Genetic Studies on the Sea Urchin Embryogenesis and Skeletogenesis (성게의 발생과 뼈대형성의 유전학적 연구)

  • Lee, Youn-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.6 no.4
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    • pp.265-273
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    • 2001
  • The sea urchin has been used as sea food in many countries. This species has also been an important organism of embryological studies for more than a century. In recent years, sea urchin embryos are being used as testing materials for toxicity of pollutants and toxins. Usefulness of sea urchin embryos as experimental models comes from the easiness in obtaining sea urchin samples and a lot of gametes, in rearing embryos in the laboratory, in observing the cellular movement and organ formation during the embryogenesis and in manipulating blastomeres and genetic maferials. The sea urchin in itself is a key organism for the understanding of deuterostome evolution from the protostomes and of indirect development of marine invertebrates which undergo the planktotrophic larval stage. A fertilized sea urchin egg goes through rapid cleavage and becomes a 60 cell embryo 7hr after fertilization. It then develops into a morula, a blastula, a gastrula and finally a pluteus larva approximately 70 hr after fertilization. At the 60 cell stage, the embryo comprises of five territories that express territory-speciflc genes and later form different organs. Micromeres at the vegetal pole ingress into the blastoceol and become the primary mesenchyme cells(PMCs). PMCs express genes involved in skeletogenesis such as SM30, SM37, SM50, PM27, msp130. Among the genes, SM37 and SM50 are considered to be members of a gene family which is characterized by early blastula expression, Glycine-Proline-Glutamine rich repeat structures and spicule matrix forming basic proteins. Genetic studies on the sea urchin embryos help understand the molecular basis of indirect development of marine invertebrates and also of the biomineralization common to the animal kingdom.

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Inhibition of Quorum Sensing and Biofilm Formation by Synthetic Quorum Signal Analogues in Pseudomonas aeruginosa (합성된 쿼럼 신호 유사 물질에 의한 녹농균 쿼럼 센싱 및 생물막 형성의 제어)

  • Kim, Soo-Kyoung;Kim, Cheol-Jin;Yoon, Je-Yong;Lee, Joon-Hee
    • Microbiology and Biotechnology Letters
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    • v.39 no.1
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    • pp.29-36
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    • 2011
  • Pseudomonas aeruginosa is an opportunistic pathogen that causes various infections on urinary track, cornea, respiratory track, and burn wound site, and mainly relies on quorum sensing (QS) for its virulence. To control the infectivity of P. aeruginosa, we previously synthesized the structural analogues of a major QS signal, N-3-oxododecanoyl homoserine lactone (3OC12-HSL) to use as a QS inhibitor. Two of them (5b and 5f) had been confirmed to have an inhibitory effect on LasR, a major QS signal receptor of P. aeruginosa in the screening by the recombinant Escherichia coli reporter. To further evaluate these compounds, we tested their efficacy to control the QS and virulence of P. aeruginosa. Unlike the result from E. coli reporter, both 5b and 5f failed to affect the LasR activity in P. aeruginosa, but instead they selectively affected the activity of QscR, another 3OC12-HSL receptor of P. aeruginosa. Interestingly, their effect on QscR was complex and opposite to what we obtained with E. coli system. Both 5b and 5f enhanced the QscR activity at the low concentration range (< 10 ${\mu}m$), but high concentration of 5f (${\approx}$1 mM) strongly inhibited QscR. While 5b and 5f didn't affect the production of proteases, the key virulence factor, they significantly reduced the biofilm formation that is important in mediating chronic infections. Especially, 5f inhibited the initial attachment of P. aeruginosa, rather than the biofilm maturation. Based on our results, we suggest that 5f can be applied for an anti-biofilm agent without increasing virulence of P. aeruginosa.

Transport Properties of CO2 and CH4 using Poly(ether-block-amide)/GPTMS Hybird Membranes (Poly(ether-block-amide)/GPTMS 하이브리드 분리막을 이용한 이산화탄소와 메탄의 투과특성)

  • Lee, Keun Chul;Kim, Hyunjoon
    • Korean Chemical Engineering Research
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    • v.54 no.5
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    • pp.653-658
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    • 2016
  • Poly(ether-block-amide)(PEBAX$_{(R)}$) resin is a thermoplastic elastomer combining linear chains of hard-rigid polyamide block interspaced soft-flexible polyether block. It was believed that the hard polyamide block provides the mechanical strength and permselectivity, whereas gas transport occurs primarily through the soft polyether block. The objective of this work was to investigate the gas permeation properties of carbon dioxide and methane for PEBAX$^{(R)}$-1657 membrane, and compare with those obtained for other grade of pure PEBAX$^{(R)}$, PEBAX$^{(R)}$-2533 and PEBAX$^{(R)}$ based hybrid membranes. The hybrid membranes based PEBAX$^{(R)}$ were obtained by a sol-gel process using GPTMS ((3-glycidoxypropyl) trimethoxysilane) as the only inorganic precursor. Molecular structure and morphology of membrane were analyzed by $^{29}Si$-NMR, DSC and SEM. PEBAX$_{(R)}$-2533 membrane exhibited higher gas permeability coefficients than PEBAX$^{(R)}$-1657 membrane. This was explained by the increase of chain mobility. In contrast, ideal separation factor of $CO_2/CH_4$ for PEBAX$^{(R)}$-1657 membrane was higher than PEBAX$^{(R)}$-2533 membrane. It was explained by the decrease of diffusion selectivity caused by increase of chain mobility. For PEBAX$^{(R)}$/GPTMS hybrid membrane, gas permeability coefficients were decreased with reaction time. Gas permeability coefficient of $CH_4$ was more significantly decreased than $CO_2$. It can be explained by the reduction of chain mobility caused by the sol-gel process, and strong affinity of PEO segment with $CO_2$. Comparing with pure PEBAX$^{(R)}$-1657 membrane, ideal separation factor of $CO_2/CH_4$ for PEBAX$^{(R)}$/GPTMS hybrid membrane has decreased to 4.5%, and gas permeability coefficient of $CO_2$ has increased 3.5 times.

Pharmacological Comparison of Timosaponin A III on the 5-beta Reductase and Androgen Receptor via In Silico Molecular Docking Approach (In silico 약리학적 분석을 통한 티모사포닌 A III의 5-베타 리덕타아제 단백질 및 안드로겐 수용체 단백질 활성 부위에 대한 결합 친화도 비교 연구)

  • Kim, Dong-Chan
    • Journal of Life Science
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    • v.28 no.3
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    • pp.307-313
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    • 2018
  • Alopecia cause psychological stress due to their effect on appearance. Thus, the global market size of the alopecia treatment products are growing quickly. Timosaponin A III is the well known active ingredient of Anemarrhenae Rhizoma. In this study, we investigated and compared the binding affinity of timosaponin A III with finasteride (5-beta reductase antagonist) and minoxidil (androgen receptor antagonist) on the target protein active site by in silico computational docking studies. The three dimensional crystallographic structure of 5-beta reductase (PDB ID : 3G1R) and androgen receptor (PDB ID: 4K7A) was obtained from PDB database. In silico computational autodocking analysis was performed using PyRx, Autodock Vina, Discovery Studio Version 4.5, and NX-QuickPharm option based on scoring functions. The timosaponin A III showed optimum binding affinity (docking energy) with 5-beta reductase as -12.20 kcal/mol as compared to the finasteride (-11.70 kcal/mol) and with androgen receptor as -9.00 kcal/mol as compared to the minoxidil (-7.40 kcal/mol). The centroid X, Y, Z grid position of the timosaponin A III on the 5-beta reductase was similar (overlap) to the finasteride, but the X, Y, Z centroid grid of the timosaponin A III on the androgen receptor was significantly far from the minoxidil centroid position. These results significantly indicated that timosaponin A III could be more potent antagonist to the 5-beta reductase and androgen receptor. Therefore, the extract of Anemarrhenae Rhizoma or timosaponin A III containing biomaterials can substitute the finasteride and minoxidil and can be applied to the alopecia protecting product and related industrial fields.

The Preparation and Physicochemical Properties of Dipalmitoylphosphatidylcholine/Cholesterol/Fluorinated Surfactant Vesicle Incorporated Fatty Acid Salt (불소화지방산염 첨가에 의한 디팔미토일포스파티딜콜린/콜레스테롤/불소화계면활성제 베지클의 제조와 물성 측정 연구)

  • Park, Young Ju;Kwon, Kyung Ok;Kim, Myung Ja
    • Applied Chemistry for Engineering
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    • v.9 no.3
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    • pp.457-461
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    • 1998
  • The vesicle system of DPPC(dipalmitoylphosphaticylcholine)/Chol(Cholesterol) has been modified by incorporating various mole fractions of flourinated surfactant($C_8F_{17}(CH_2)_2OCO-CH_2CH(SO_3Na)COO(CH_2)_2C_8F_{17}$. Sodium bis(1H,1H,2H,2H-heptadecaflurododecyl)-2-sulfosuccinate, FS)/fluorinated fatty acid salt ($C_7F_{15}COONH_4$, ammoniumpentadecaflurooctyrate, FFS), and their physicochemical properties have been investigated in an attempt to enhance the stability of phospholipid vesicle system. The ${\zeta}$-potential measurement by use of Zetamaster sub-micron Particle Electrophoresis Analyzer (Malvern Co.) showed that a charged homogeneous DPPC/Chol/FS vesicle has been formed owing to the incorporated FFS effect on the membrane, playing a role as a cosurfactant in the bilayer between DPPC and FS components. With increase in the concentration of FFS, it was found that the particle size and also surface charge of the DPPC/Chol/FS vesicle decreased. The stability of DPPC/Chol/FS/FFS liposome was found to be enhanced significantly compared to that of DPPC/Chol/FS according to the dispersity change as a function of time. The release rate of dye molecule of Methylene Blue from the DPPC/Chol/FS/FFS vesicle was determined to be slower than that of DPPC/Chol/FS system, and it may be attributed to the increase in microviscosity of the hydrophobic region in the bilayer. The affinfinity of DPPC/Chol/FS/FFS vesicles to albumin was found to be slightly lowered compared to that of DPPC/Chol/FS. Based on these findings, it was confirmed that a more stable and homogeneous vesicle system of DPPC/Chol/FS could be prepared by addition of FFS, acting as a cosurfactant in the aggregate formation.

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