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  1 / 73 MEDLINE  
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PMID:29020603
Autor:Inan C; Muratoglu H; Arif BM; Demirbag Z
Dirección:Department of Biology, Faculty of Sciences, Karadeniz Technical University, Trabzon, Turkey; Department of Molecular Biology and Genetics, Faculty of Sciences, Karadeniz Technical University, Trabzon, Turkey.
Título:Transcriptional analysis of the putative glycosyltransferase gene (amv248) of the Amsacta moorei entomopoxvirus.
Fuente:Virus Res; 243:25-30, 2018 01 02.
ISSN:1872-7492
País de publicación:Netherlands
Idioma:eng
Resumen:Amsacta moorei entomopoxvirus (AMEV), the most studied member of the genus Betaentomopoxvirus, was initially isolated from Red Hairy caterpillar larvae, Amsacta moorei. According to genome sequence and previous studies it was shown that amv248 encodes a putative glycosyltransferase that is the only conserved attachment protein in betaentomopoxviruses. Transcriptional analysis of the amv248 gene by RT-PCR and qPCR showed that transcription starts at 6h post infection (hpi). Also, transcription was not affected by a DNA replication inhibitor but was severely curtailed by a protein synthesis inhibitor. These results indicate that amv248 belongs to the intermediate class of gene expression. 5' and 3' untranslated regions analysis revealed that transcription initiates at position -126 relative to the translational start site, and ends between 50 and 83 bases after the stop codon. To narrow down the size and location of the gene's promoter, the upstream region as well as several different sized deletions thereof were generated and cloned upstream of a luciferase reporter gene. The constructs were used to measure the Firefly and Renilla luciferase activities in dual assays. The results showed that luciferase activity decreased when bases -198 to -235 of amv248 upstream region were missing. Sequence analysis among the intermediate gene promoters of AMEV showed that TTTAT(T/A)TT(T/A) TTA is possibly a common motif, however, further investigations are needed to confirm this conclusion.
Tipo de publicación:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
Nombre de substancia:0 (Viral Proteins); EC 2.4.- (Glycosyltransferases)


  2 / 73 MEDLINE  
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PMID:26820433
Autor:Muratoglu H; Nalcacioglu R; Arif BM; Demirbag Z
Dirección:Karadeniz Technical University, Faculty of Sciences, Department of Molecular Biology and Genetic, 61080 Trabzon, Turkey.
Título:Genome-wide analysis of differential mRNA expression of Amsacta moorei entomopoxvirus, mediated by the gene encoding a viral protein kinase (AMV197).
Fuente:Virus Res; 215:25-36, 2016 Apr 02.
ISSN:1872-7492
País de publicación:Netherlands
Idioma:eng
Resumen:Insect-born entomopoxviruses (Fam: Poxviridae) are potentially important bio-pesticide against insect pests and expression vectors as well as vectors for transient human gene therapies including recombinant viral vaccines. For these reasons, it is necessary to understand the regulatory genes functions to improve its biotechnological potential. Here, we focused on the characterization of serine/threonine (Ser/Thr; ORF AMV197) protein kinase gene from the Amsacta moorei entomopoxvirus (AMEV), the type species of the genus Betaentomopoxvirus. Transcription of the parental and an amv197-null recombinant AMEV was compared by whole-genome gene expression microarray analysis. Blast2GO analysis reflected a broad diversity of upregulated and downregulated genes. Results showed that expression levels of 102 genes (45%) out of 226 tested genes changed significantly in the recombinant AMEV infected cells. Of these transcripts, 72 (70.58%) were upregulated and 30 (29.41%) were downregulated throughout the infection period. Genes involved in DNA repair, replication and nucleotide metabolism, transcription and RNA modification, and protein modification were mostly upregulated at different times in cells infected with the recombinant virus. Furthermore, transcription of all studied cellular genes including metabolism of apoptosis (Nedd2-like caspase, hemolin and elongation factor-1 alpha (ef1a) gene) was downregulated in the absence of amv197. Quantitative real time reverse transcription-PCR confirmed viral transcriptional changes obtained by microarray. The results of this study indicated that the product of amv197 appears to affect the transcriptional regulation of most viral and many cellular genes. Further investigations are, however, needed to narrow down the role of AMV197 throughout the infection process.
Tipo de publicación:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
Nombre de substancia:0 (RNA, Messenger); EC 2.7.- (Protein Kinases)


  3 / 73 MEDLINE  
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PMID:26499185
Autor:Nakai M; Kinjo H; Takatsuka J; Shiotsuki T; Kamita SG; Kunimi Y
Dirección:1​ Tokyo University of Agriculture and Technology, Saiwai, , Fuchu, Tokyo 183-8509, Japan.
Título:Entomopoxvirus infection induces changes in both juvenile hormone and ecdysteroid levels in larval Mythimna separata.
Fuente:J Gen Virol; 97(1):225-32, 2016 Jan.
ISSN:1465-2099
País de publicación:England
Idioma:eng
Resumen:Insect viruses are among the most important pathogens of lepidopteran insects. Virus-infected larvae often show developmental defects including a prolonged larval period and a failure to pupate, but the mechanisms by which insect viruses regulate host development need further investigation. In this study, the regulation of host endocrinology by a lepidopteran entomopoxvirus (EPV), Mythimna separata EPV (MySEV), was examined. When fourth instar M. separata were inoculated with MySEV occlusion bodies, pupation was prevented and the insects died during the final (sixth) larval instar. Liquid chromatography-MS analysis revealed that juvenile hormone (JH) titres in the haemolymph of MySEV-infected sixth instars were higher than those in mock-infected larvae. JH esterase (JHE) activity was also examined by kinetic assay using a colorimetric substrate. The level of JHE activity in the haemolymph of MySEV-infected larvae was generally lower than that found in mock-infected larvae. In contrast, ecdysteroid titre in the haemolymph of final-instar MySEV-infected larvae was lower than that found in mock-infected larvae when measured by radioimmunoassay. A statistically significant difference in the release of ecdysteroids from prothoracic glands (PGs) that were dissected from MySEV- or mock-infected sixth instar Day 3 larvae was not found following prothoracicotropic hormone (PTTH) exposure. Our results indicate that the release of ecdysteroids was reduced not by infection of the PGs by MySEV, but by reduced PTTH production from the brain. Taken together our study suggests that EPVs retard host development by both reducing ecdysone titre and maintaining status quo levels of JH by preventing its metabolism.
Tipo de publicación:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
Nombre de substancia:0 (Ecdysteroids); 0 (Juvenile Hormones); EC 3.1.- (Esterases)


  4 / 73 MEDLINE  
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PMID:26204184
Autor:Lukásová K; Holusa J
Título:Comparison of pathogens infection level in Ips typographus (Coleoptera: Curculionidae) beetles sampled in pheromone traps and at place of overwintering.
Fuente:Acta Parasitol; 60(3):462-5, 2015 Sep.
ISSN:1896-1851
País de publicación:Poland
Idioma:eng
Resumen:The importance of pathogens in the population dynamics of Ips typographus remains a subject of ongoing debate. The main objective of our experiment was to compare the pathogen infection levels of individuals overwintering in bark with the levels of individuals from the same population captured with pheromone traps and thereby to determine primary answers as to whether it can be confirmed that pathogenic organisms affect the flight ability of bark beetles or their ability to leave their places of overwintering. A total of 402 I. typographus individuals were analyzed at a study location under limited management. Three pathogens were confirmed to be present: the gregarine Gregarina typographi, the virus ItEPV, and the microsporidium Nosema typographi. Infection levels of Gregarina typographi and ItEPV were the same in beetles collected at places of overwintering and in those beetles collected in pheromone traps within the immediate vicinity. As these pathogens infect the host's intestine, the tendency to leave the places of overwintering is apparently not diminished. A similar analysis and comparison of pathogens located in the fat body might bring different results, as our study only detected N. typographi in a single dissected adult spruce bark beetle.
Tipo de publicación:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T


  5 / 73 MEDLINE  
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PMID:25871928
Autor:Thézé J; Takatsuka J; Nakai M; Arif B; Herniou EA
Dirección:Institut de Recherche sur la Biologie de l'Insecte, CNRS UMR 7261, Université François-Rabelais, UFR Sciences et Techniques, 37200 Tours, France. theze.julien@gmail.com.
Título:Gene acquisition convergence between entomopoxviruses and baculoviruses.
Fuente:Viruses; 7(4):1960-74, 2015 Apr 13.
ISSN:1999-4915
País de publicación:Switzerland
Idioma:eng
Resumen:Organisms from diverse phylogenetic origins can thrive within the same ecological niches. They might be induced to evolve convergent adaptations in response to a similar landscape of selective pressures. Their genomes should bear the signature of this process. The study of unrelated virus lineages infecting the same host panels guarantees a clear identification of phyletically independent convergent adaptation. Here, we investigate the evolutionary history of genes in the accessory genome shared by unrelated insect large dsDNA viruses: the entomopoxviruses (EPVs, Poxviridae) and the baculoviruses (BVs). EPVs and BVs have overlapping ecological niches and have independently evolved similar infection processes. They are, in theory, subjected to the same selective pressures from their host's immune responses. Their accessory genomes might, therefore, bear analogous genomic signatures of convergent adaption and could point out key genomic mechanisms of adaptation hitherto undetected in viruses. We uncovered 32 homologous, yet independent acquisitions of genes originating from insect hosts, different eukaryotes, bacteria and viruses. We showed different evolutionary levels of gene acquisition convergence in these viruses, underlining a continuous evolutionary process. We found both recent and ancient gene acquisitions possibly involved to the adaptation to both specific and distantly related hosts. Multidirectional and multipartite gene exchange networks appear to constantly drive exogenous gene assimilations, bringing key adaptive innovations and shaping the life histories of large DNA viruses. This evolutionary process might lead to genome level adaptive convergence.
Tipo de publicación:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
Nombre de substancia:0 (DNA, Viral)


  6 / 73 MEDLINE  
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PMID:25591507
Autor:Özsahin E; Sezen K; Demirbag Z
Dirección:Department of Biology, Faculty of Science, Karadeniz Technical University, Trabzon, Turkey.
Título:Amsacta moorei entomopoxvirus encodes a functional esterase (amv133) with protease activity.
Fuente:Intervirology; 58(1):41-8, 2015.
ISSN:1423-0100
País de publicación:Switzerland
Idioma:eng
Resumen:OBJECTIVES: Lipolytic genes have been investigated in several viral genomes, and some of them show enzyme activity which can be used for various functions including the production of DNA replication metabolites, rescue from endosomes, and membrane fusion. Amsacta moorei entomopoxvirus (AMEV) replicates in nearly the entire insect body, especially in the adipose tissue. One of the open reading frames (ORFs) in the AMEV genome, amv133, encodes a putative lipase enzyme. In this study, we therefore investigate the enzyme activity of amv133. METHODS: amv133 was aligned with known lipase genes and their homologs in entomopoxviruses. Expressed proteins were partially purified and assayed for lipase, esterase and protease. RESULTS: We found that amv133 contains all the domains required for a functional lipase enzyme and that it shows a significant similarity with homologs in other entomopoxviruses. Since there is a similarity of the catalytic triad between lipases and serine proteases, we also investigated the protease activity of amv133. Lipase, esterase and protease assays showed that amv133 encodes a functional esterase enzyme with protease activity. CONCLUSION: The current data show that amv133 is a conserved gene in all entomopoxvirus genomes sequenced so far and might contribute greatly to degrading the lipids or proteins and hence improve the virus infection.
Tipo de publicación:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
Nombre de substancia:0 (Viral Proteins); EC 3.1.- (Esterases); EC 3.1.1.3 (Lipase); EC 3.4.- (Peptide Hydrolases)


  7 / 73 MEDLINE  
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PMID:24796553
Autor:Ozsahin E; Sezen K; Demirbag Z
Dirección:Department of Biology, Faculty of Science, Karadeniz Technical University, Trabzon, Turkey.
Título:Transcriptional analysis of ORF amv133 of Amsacta moorei entomopoxvirus.
Fuente:Arch Virol; 159(10):2541-7, 2014 Oct.
ISSN:1432-8798
País de publicación:Austria
Idioma:eng
Resumen:The open reading frame (ORF) amv133 of Amsacta moorei entomopoxvirus, encodes a putative lipase gene. Its temporal expression pattern was characterized by RT-PCR and found to start at 6 h postinfection (h p.i.) and reach a maximum level at 48 h p.i. While the ORF has a late promoter motif, the inhibition of viral DNA synthesis by Ara-C failed to inhibit transcription, but a general inhibitor of protein synthesis prevented its transcription, indicating that amv133 is an intermediate gene. 5'-RACE analysis showed that transcription was initiated at position -77 relative to the translational start site. To determine the size of the promoter, several truncations were generated and cloned upstream of the firefly luciferase reporter gene. The resulting constructs were tested in a dual assay. A fragment that contained 115 bp relative to the transcription start site exhibited optimum promoter length.
Tipo de publicación:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
Nombre de substancia:0 (5' Untranslated Regions); 0 (Antiviral Agents); 0 (DNA, Viral); 0 (Viral Proteins); 04079A1RDZ (Cytarabine); EC 3.1.1.3 (Lipase)


  8 / 73 MEDLINE  
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PMID:24606687
Autor:Mitsuhashi W; Miyamoto K; Wada S
Dirección:National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8634, Japan. Electronic address: mitsuhas@affrc.go.jp.
Título:The complete genome sequence of the Alphaentomopoxvirus Anomala cuprea entomopoxvirus, including its terminal hairpin loop sequences, suggests a potentially unique mode of apoptosis inhibition and mode of DNA replication.
Fuente:Virology; 452-453:95-116, 2014 Mar.
ISSN:1096-0341
País de publicación:United States
Idioma:eng
Resumen:Complete genome sequence of Anomala cuprea entomopoxvirus, which belongs to the genus Alphaentomopoxvirus, including its terminal hairpin loop sequences, is reported. This is the first genome sequence of Alphaentomopoxvirus reported, and hairpin loops in entomopoxviruses have not previously been sequenced. The genome is 245,717 bp, which is smaller than had previously been estimated for Alphaentomopoxvirus. The inverted terminal repeats are quite long, and experimental results suggest that one genome molecule has one type of hairpin at one end and another type at the other end. The genome contains unexpected ORFs, e.g., that for the ubiquitin-conjugating enzyme E2 of eukaryotes. The BIR and RING domains found in a single ORF for an inhibitor of apoptosis in baculoviruses and entomopoxviruses occurred in two different, widely separated ORFs. Furthermore, an ORF in the genome contains a serpin domain that was previously found in vertebrate poxviruses for apoptosis inhibition but not in insect viruses.
Tipo de publicación:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
Nombre de substancia:0 (DNA, Viral); 0 (Viral Proteins)


  9 / 73 MEDLINE  
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PMID:24603041
Autor:Takatsuka J; Nakai M
Dirección:Forestry and Forest Products Research Institute, Matsunosato, Tsukuba, Ibaraki 305-8687, Japan. Electronic address: junsan@ffpri.affrc.go.jp.
Título:Replication of Mythimna separata entomopoxvirus in High Five™ cells and the construction of a recombinant.
Fuente:J Invertebr Pathol; 118:12-7, 2014 May.
ISSN:1096-0805
País de publicación:United States
Idioma:eng
Resumen:Mythimna separata entomopoxvirus (MySEV), of the genus Betaentomopoxvirus, was found to replicate in High Five™ cells. The infected cells produced many occlusion bodies and were hypertrophied but did not lyse. Following infection at a multiplicity of infection of 0.1, titers of extracellular virus reached a plateau 3-4days post infection at 25°C and were estimated at ca. 3×10(5) plaque-forming units per ml in TC-100 or TMN-FH media, both of which contained fetal bovine serum (FBS). Serum free medium, Express Five® SFM, also supported virus replication in High Five™ cells, but the titers were approximately one-tenth of those grown in TC-100 or TMN-FH media containing FBS. Using High Five™ cells, a recombinant MySEV was successfully constructed using homologous recombination. This study opens an avenue to the evaluation of entomopoxvirus gene functions using reverse genetic approaches with in vitro and in vivo hosts.
Tipo de publicación:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T


  10 / 73 MEDLINE  
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PMID:23424042
Autor:Mitsuhashi W; Asano S; Miyamoto K; Wada S
Dirección:National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan.
Título:Further research on the biological function of inclusion bodies of Anomala cuprea entomopoxvirus, with special reference to the effect on the insecticidal activity of a Bacillus thuringiensis formulation.
Fuente:Pest Manag Sci; 70(1):46-54, 2014 Jan.
ISSN:1526-4998
País de publicación:England
Idioma:eng
Resumen:BACKGROUND: Entomopoxviruses (EVs) form two types of inclusion body: spheroids, which contain virions, and spindles, which do not. The authors tested whether the spindles from a coleopteran EV, Anomala cuprea EV (ACEV), enhanced the insecticidal activity of a commercial Bacillus thuringiensis (Bt) formulation and the susceptibility of scarabaeid pest species in Japan to the virus's spheroids, to assess whether ACEV inclusion bodies are potential biological control agents for pest insects. RESULTS: Peroral inoculation with both ACEV spindles and the Bt toxin only or the complete Bt formulation shortened the survival and increased the mortality of treated insects compared with those of insects inoculated with Bt without the spindles (8-38 h of decrease in LT50 values among assays). ACEV showed high infectivity to a major scarabaeid pest species in Japanese sugar cane fields. CONCLUSION: The results suggest that spindles or the constituent protein fusolin can be used as a coagent with Bt formulations, and that fusolin coexpression with a Bt toxin in crops might improve the insecticidal efficacy. In addition, the spheroids are potential biocontrol agents for some scarabaeid pests that are not easy to control because of their underground habitation.
Tipo de publicación:EVALUATION STUDIES; JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
Nombre de substancia:0 (Bacterial Proteins); 0 (Endotoxins); 0 (Hemolysin Proteins); 0 (insecticidal crystal protein, Bacillus Thuringiensis)



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