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[PMID]:29385169
[Au] Autor:Bissa M; Forlani G; Zanotto C; Tosi G; De Giuli Morghen C; Accolla RS; Radaelli A
[Ad] Endereço:Department of Pharmacological and Biomolecular Sciences, University of Milan, via Balzaretti 9, Milan, Italy.
[Ti] Título:Fowlpoxvirus recombinants coding for the CIITA gene increase the expression of endogenous MHC-II and Fowlpox Gag/Pro and Env SIV transgenes.
[So] Source:PLoS One;13(1):e0190869, 2018.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:A complete eradication of an HIV infection has never been achieved by vaccination and the search for new immunogens that can induce long-lasting protective responses is ongoing. Avipoxvirus recombinants are host-restricted for replication to avian species and they do not have the undesired side effects induced by vaccinia recombinants. In particular, Fowlpox (FP) recombinants can express transgenes over long periods and can induce protective immunity in mammals, mainly due to CD4-dependent CD8+ T cells. In this context, the class II transactivator (CIITA) has a pivotal role in triggering the adaptive immune response through induction of the expression of class-II major histocompatibility complex molecule (MHC-II), that can present antigens to CD4+ T helper cells. Here, we report on construction of novel FPgp and FPenv recombinants that express the highly immunogenic SIV Gag-pro and Env structural antigens. Several FP-based recombinants, with single or dual genes, were also developed that express CIITA, driven from H6 or SP promoters. These recombinants were used to infect CEF and Vero cells in vitro and determine transgene expression, which was evaluated by real-time PCR and Western blotting. Subcellular localisation of the different proteins was evaluated by confocal microscopy, whereas HLA-DR or MHC-II expression was measured by flow cytometry. Fowlpox recombinants were also used to infect syngeneic T/SA tumour cells, then injected into Balb/c mice to elicit MHC-II immune response and define the presentation of the SIV transgene products in the presence or absence of FPCIITA. Antibodies to Env were measured by ELISA. Our data show that the H6 promoter was more efficient than SP to drive CIITA expression and that CIITA can enhance the levels of the gag/pro and env gene products only when infection is performed by FP single recombinants. Also, CIITA expression is higher when carried by FP single recombinants than when combined with FPgp or FPenv constructs and can induce HLA-DR cell surface expression. However, in-vivo experiments did not show any significant increase in the humoral response. As CIITA already proved to elicit immunogenicity by improving antigen presentation, further in-vivo experiments should be performed to increase the immune responses. The use of prime/boost immunisation protocols and the oral administration route of the recombinants may enhance the immunogenicity of Env peptides presented by MHC-II and provide CD4+ T-cell stimulation.
[Mh] Termos MeSH primário: Genes Virais
Complexo Principal de Histocompatibilidade/genética
Proteínas Nucleares/genética
Poxviridae/genética
Recombinação Genética
Vírus da Imunodeficiência Símia/genética
Transativadores/genética
Transgenes
[Mh] Termos MeSH secundário: Vacinas contra a AIDS/genética
Vacinas contra a AIDS/imunologia
Animais
Western Blotting
Linfócitos T CD4-Positivos/imunologia
Linfócitos T CD8-Positivos/imunologia
Linhagem Celular
Embrião de Galinha
Ensaio de Imunoadsorção Enzimática
Infecções por HIV/imunologia
Infecções por HIV/prevenção & controle
Seres Humanos
Camundongos
Camundongos Endogâmicos BALB C
Microscopia Confocal
Regiões Promotoras Genéticas
Reação em Cadeia da Polimerase em Tempo Real
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (AIDS Vaccines); 0 (MHC class II transactivator protein); 0 (Nuclear Proteins); 0 (Trans-Activators)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180221
[Lr] Data última revisão:
180221
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180201
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0190869


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[PMID]:28976983
[Au] Autor:Stading B; Ellison JA; Carson WC; Satheshkumar PS; Rocke TE; Osorio JE
[Ad] Endereço:Department of Pathobiological Sciences, University of Wisconsin - Madison, Madison, Wisconsin, United States of America.
[Ti] Título:Protection of bats (Eptesicus fuscus) against rabies following topical or oronasal exposure to a recombinant raccoon poxvirus vaccine.
[So] Source:PLoS Negl Trop Dis;11(10):e0005958, 2017 Oct.
[Is] ISSN:1935-2735
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Rabies is an ancient neglected tropical disease that causes tens of thousands of human deaths and millions of cattle deaths annually. In order to develop a new vaccine for potential use in bats, a reservoir of rabies infection for humans and animals alike, an in silico antigen designer tool was used to create a mosaic glycoprotein (MoG) gene using available sequences from the rabies Phylogroup I glycoprotein. This sequence, which represents strains more likely to occur in bats, was cloned into raccoonpox virus (RCN) and the efficacy of this novel RCN-MoG vaccine was compared to RCN-G that expresses the glycoprotein gene from CVS-11 rabies or luciferase (RCN-luc, negative control) in mice and big brown bats (Eptesicus fuscus). Mice vaccinated and boosted intradermally with 1 x 107 plaque forming units (PFU) of each RCN-rabies vaccine construct developed neutralizing antibodies and survived at significantly higher rates than controls. No significant difference in antibody titers or survival was noted between rabies-vaccinated groups. Bats were vaccinated either oronasally (RCN-G, RCN-MoG) with 5x107 PFU or by topical application in glycerin jelly (RCN-MoG, dose 2x108 PFU), boosted (same dose and route) at 46 days post vaccination (dpv), and then challenged with wild-type big brown variant RABV at 65 dpv. Prior to challenge, 90% of RCN-G and 75% of RCN-MoG oronasally vaccinated bats had detectable levels of serum rabies neutralizing antibodies. Bats from the RCN-luc and topically vaccinated RCN-MoG groups did not have measurable antibody responses. The RCN-rabies constructs were highly protective and not significantly different from each other. RCN-MoG provided 100% protection (n = 9) when delivered oronasally and 83% protection (n = 6) when delivered topically; protection provided by the RCN-G construct was 70% (n = 10). All rabies-vaccinated bats survived at a significantly (P ≤ 0.02) higher rate than control bats (12%; n = 8). We have demonstrated the efficacy of a novel, in silico designed rabies MoG antigen that conferred protection from rabies challenge in mice and big brown bats in laboratory studies. With further development, topical or oronasal administration of the RCN-MoG vaccine could potentially mitigate rabies in wild bat populations, reducing spillover of this deadly disease into humans, domestic mammals, and other wildlife.
[Mh] Termos MeSH primário: Quirópteros
Poxviridae/imunologia
Raiva/veterinária
Vacinas Virais
[Mh] Termos MeSH secundário: Animais
Linhagem Celular
Cricetinae
Feminino
Masculino
Camundongos
Raiva/mortalidade
Raiva/prevenção & controle
Vacinas Sintéticas
Vacinas Virais/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Vaccines, Synthetic); 0 (Viral Vaccines)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171103
[Lr] Data última revisão:
171103
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171005
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pntd.0005958


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[PMID]:28895845
[Au] Autor:Athanasopoulos T; Munye MM; Yáñez-Muñoz RJ
[Ad] Endereço:Cell and Gene Therapy Discovery Research, Platform Technology and Sciences, GSK Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertfordshire SG1 2NY, UK.
[Ti] Título:Nonintegrating Gene Therapy Vectors.
[So] Source:Hematol Oncol Clin North Am;31(5):753-770, 2017 Oct.
[Is] ISSN:1558-1977
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Gene delivery vectors that do not rely on host cell genome integration offer several advantages for gene transfer, chiefly the avoidance of insertional mutagenesis and position effect variegation. However, unless engineered for replication and segregation, nonintegrating vectors will dilute progressively in proliferating cells, and are not exempt of epigenetic effects. This article provides an overview of the main nonintegrating viral (adenoviral, adeno-associated viral, integration-deficient retro-lentiviral, poxviral), and nonviral (plasmid vectors, artificial chromosomes) vectors used for preclinical and clinical cell and gene therapy applications. Particular emphasis is placed on their use in hematologic disease.
[Mh] Termos MeSH primário: Terapia Genética
Vetores Genéticos/genética
[Mh] Termos MeSH secundário: Adenoviridae/genética
Animais
Ensaios Clínicos como Assunto/história
Dependovirus/genética
Edição de Genes
Expressão Gênica
Técnicas de Transferência de Genes
Terapia Genética/efeitos adversos
Terapia Genética/história
Terapia Genética/métodos
Vetores Genéticos/classificação
História do Século XX
História do Século XXI
Seres Humanos
Plasmídeos/genética
Poxviridae/genética
Transdução Genética
[Pt] Tipo de publicação:HISTORICAL ARTICLE; JOURNAL ARTICLE; REVIEW
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171009
[Lr] Data última revisão:
171009
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170913
[St] Status:MEDLINE


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[PMID]:28604109
[Au] Autor:Asad AS; Moreno Ayala MA; Gottardo MF; Zuccato C; Nicola Candia AJ; Zanetti FA; Seilicovich A; Candolfi M
[Ad] Endereço:a Departamento de Biología Celular e Histología, Facultad de Medicina , Universidad de Buenos Aires , Buenos Aires , Argentina.
[Ti] Título:Viral gene therapy for breast cancer: progress and challenges.
[So] Source:Expert Opin Biol Ther;17(8):945-959, 2017 Aug.
[Is] ISSN:1744-7682
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:INTRODUCTION: Breast cancer is the most common cancer in women all over the world. Furthermore, up to one third of breast tumors develop metastases that are resistant to standard therapies. Gene therapeutic strategies have been developed in order to specifically target cancer cells either directly or through the stimulation of antitumor immunity. Areas covered: This review describes the therapeutic strategies that are currently under development to treat this disease using engineered viral vectors including: adenovirus, adeno-associated virus, lentivirus, poxvirus, reovirus, baculovirus, herpesvirus and oncolytic viruses. Advantages and disadvantages of these multiple gene therapy platforms are discussed in detail. Expert opinion: Metastatic breast cancer is a perfect candidate for gene therapy approaches due to the presence of several tumor antigens and the aberrant expression of many molecular pathways. Oncolytic vectors are able to attack tumor cells while sparing normal cells and their activity is often enhanced by the administration of chemotherapy. However, more efforts are needed in order to reduce toxicity and to achieve better transduction efficiency. Improved preclinical models and a more critical patient selection for clinical trials, along with advances in gene therapy regulations, will surely facilitate the evolution of gene therapy for the treatment of metastatic breast cancer.
[Mh] Termos MeSH primário: Neoplasias da Mama/terapia
Terapia Viral Oncolítica
[Mh] Termos MeSH secundário: Adenoviridae/genética
Feminino
Vetores Genéticos/genética
Vetores Genéticos/metabolismo
Seres Humanos
Lentivirus/genética
Células Mesenquimais Estromais/citologia
Células Mesenquimais Estromais/metabolismo
Células Mesenquimais Estromais/virologia
Células-Tronco Neurais/citologia
Células-Tronco Neurais/metabolismo
Células-Tronco Neurais/virologia
Vírus Oncolíticos/genética
Poxviridae/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171113
[Lr] Data última revisão:
171113
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170613
[St] Status:MEDLINE
[do] DOI:10.1080/14712598.2017.1338684


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[PMID]:28335511
[Au] Autor:McCormick C; Grandvaux N
[Ad] Endereço:Department of Microbiology and Immunology, Dalhousie University, 5850 College Street, Halifax, NS B3H 4R2, Canada. craig.mccormick@dal.ca.
[Ti] Título:1st Workshop of the Canadian Society for Virology.
[So] Source:Viruses;9(3), 2017 Mar 20.
[Is] ISSN:1999-4915
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:The 1st Workshop of the Canadian Society for Virology (CSV2016) was a Special Workshop of the 35th Annual Meeting for the American Society for Virology, held on 18 June 2016 on the beautiful Virginia Tech campus in Blacksburg, Virginia. The workshop provided a forum for discussion of recent advances in the field, in an informal setting conducive to interaction with colleagues. CSV2016 featured two internationally-renowned Canadian keynote speakers who discussed translational virology research; American Society for Virology President Grant McFadden (then from University of Florida, now relocated to Arizona State University) who presented his studies of oncolytic poxviruses, while Matthew Miller (McMaster University) reviewed the prospects for a universal influenza vaccine. The workshop also featured a variety of trainee oral and poster presentations, and a panel discussion on the topic of the future of the CSV and virus research in Canada.
[Mh] Termos MeSH primário: Pesquisa Biomédica/tendências
Sociedades Científicas
Virologia/tendências
[Mh] Termos MeSH secundário: Canadá
Educação
Seres Humanos
Vacinas contra Influenza/imunologia
Vacinas contra Influenza/isolamento & purificação
Vírus Oncolíticos/fisiologia
Poxviridae/fisiologia
Pesquisa Médica Translacional/tendências
Virginia
[Pt] Tipo de publicação:CONGRESSES
[Nm] Nome de substância:
0 (Influenza Vaccines)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170926
[Lr] Data última revisão:
170926
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170325
[St] Status:MEDLINE


  6 / 1746 MEDLINE  
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[PMID]:28282449
[Au] Autor:Aguirre de Cárcer D; Hernáez B; Rastrojo A; Alcamí A
[Ad] Endereço:Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas (CSIC)-Universidad Autónoma de Madrid (UAM), Madrid, Spain.
[Ti] Título:Infection with diverse immune-modulating poxviruses elicits different compositional shifts in the mouse gut microbiome.
[So] Source:PLoS One;12(3):e0173697, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:It is often not possible to demonstrate causality within the context of gut microbiota dysbiosis-linked diseases. Thus, we need a better understanding of the mechanisms whereby an altered host immunophysiology shapes its resident microbiota. In this regard, immune-modulating poxvirus strains and mutants could differentially alter gut mucosal immunity in the context of a natural immune response, providing a controlled natural in vivo setting to deepen our understanding of the immune determinants of microbiome composition. This study represents a proof-of-concept that the use of an existing collection of different immune-modulating poxviruses may represent an innovative tool in gut microbiome research. To this end, 16S rRNA amplicon sequencing and RNAseq transcriptome profiling were employed as proxies for microbiota composition and gut immunophysiological status in the analysis of caecal samples from control mice and mice infected with various poxvirus types. Our results show that different poxvirus species and mutants elicit different shifts in the mice mucosa-associated microbiota and, in some instances, significant concomitant shifts in gut transcriptome profiles, thus providing an initial validation to the proposed model.
[Mh] Termos MeSH primário: Microbioma Gastrointestinal/fisiologia
Infecções por Poxviridae/imunologia
Poxviridae/patogenicidade
[Mh] Termos MeSH secundário: Animais
Vírus da Ectromelia/genética
Vírus da Ectromelia/patogenicidade
Feminino
Microbioma Gastrointestinal/imunologia
Interações Hospedeiro-Patógeno/imunologia
Camundongos Endogâmicos BALB C
Mutação
Poxviridae/genética
Poxviridae/imunologia
Infecções por Poxviridae/microbiologia
Infecções por Poxviridae/fisiopatologia
RNA Ribossômico 16S
Vírus Vaccinia/genética
Vírus Vaccinia/patogenicidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Ribosomal, 16S)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170817
[Lr] Data última revisão:
170817
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170311
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0173697


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[PMID]:28222784
[Au] Autor:Adamek M; Oschilewski A; Wohlsein P; Jung-Schroers V; Teitge F; Dawson A; Gela D; Piackova V; Kocour M; Adamek J; Bergmann SM; Steinhagen D
[Ad] Endereço:Fish Disease Research Unit, Institute for Parasitology, University of Veterinary Medicine, Bünteweg 17, 30559, Hannover, Germany. Mikolaj.Adamek@tiho-hannover.de.
[Ti] Título:Experimental infections of different carp strains with the carp edema virus (CEV) give insights into the infection biology of the virus and indicate possible solutions to problems caused by koi sleepy disease (KSD) in carp aquaculture.
[So] Source:Vet Res;48(1):12, 2017 Feb 21.
[Is] ISSN:1297-9716
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Outbreaks of koi sleepy disease (KSD) caused by carp edema virus (CEV) may seriously affect populations of farmed common carp, one of the most important fish species for global food production. The present study shows further evidence for the involvement of CEV in outbreaks of KSD among carp and koi populations: in a series of infection experiments, CEV from two different genogroups could be transmitted to several strains of naïve common carp via cohabitation with fish infected with CEV. In recipient fish, clinical signs of KSD were induced. The virus load and viral gene expression results confirm gills as the target organ for CEV replication. Gill explants also allowed for a limited virus replication in vitro. The in vivo infection experiments revealed differences in the virulence of the two CEV genogroups which were associated with infections in koi or in common carp, with higher virulence towards the same fish variety as the donor fish. When the susceptibility of different carp strains to a CEV infection and the development of KSD were experimentally investigated, Amur wild carp showed to be relatively more resistant to the infection and did not develop clinical signs for KSD. However, the resistance could not be related to a higher magnitude of type I IFN responses of affected tissues. Despite not having a mechanistic explanation for the resistance of Amur wild carp to KSD, we recommend using this carp strain in breeding programs to limit potential losses caused by CEV in aquaculture.
[Mh] Termos MeSH primário: Carpas/virologia
Doenças dos Peixes/virologia
Infecções por Poxviridae/veterinária
Poxviridae
[Mh] Termos MeSH secundário: Animais
Aquicultura/métodos
Feminino
Brânquias/virologia
Masculino
Infecções por Poxviridae/virologia
Pele/virologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170928
[Lr] Data última revisão:
170928
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170223
[St] Status:MEDLINE
[do] DOI:10.1186/s13567-017-0416-7


  8 / 1746 MEDLINE  
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[PMID]:28000080
[Au] Autor:Sobhy H
[Ad] Endereço:Department of Molecular Biology, Umeå University, 901 87, Umeå, Sweden. haithamsobhy@gmail.com.
[Ti] Título:A bioinformatics pipeline to search functional motifs within whole-proteome data: a case study of poxviruses.
[So] Source:Virus Genes;53(2):173-178, 2017 Apr.
[Is] ISSN:1572-994X
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Proteins harbor domains or short linear motifs, which facilitate their functions and interactions. Finding functional motifs in protein sequences could predict the putative cellular roles or characteristics of hypothetical proteins. In this study, we present Shetti-Motif, which is an interactive tool to (i) map UniProt and PROSITE flat files, (ii) search for multiple pre-defined consensus patterns or experimentally validated functional motifs in large datasets protein sequences (proteome-wide), (iii) search for motifs containing repeated residues (low-complexity regions, e.g., Leu-, SR-, PEST-rich motifs, etc.). As proof of principle, using this comparative proteomics pipeline, eleven proteomes encoded by member of Poxviridae family were searched against about 100 experimentally validated functional motifs. The closely related viruses and viruses infect the same host cells (e.g. vaccinia and variola viruses) show similar motif-containing proteins profile. The motifs encoded by these viruses are correlated, which explains why poxviruses are able to interact with wide range of host cells. In conclusion, this in silico analysis is useful to establish a dataset(s) or potential proteins for further investigation or compare between species.
[Mh] Termos MeSH primário: Motivos de Aminoácidos/genética
Biologia Computacional
Poxviridae/genética
Proteoma/genética
[Mh] Termos MeSH secundário: Sequência de Aminoácidos/genética
Interações Hospedeiro-Patógeno/genética
Poxviridae/patogenicidade
Domínios Proteicos/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Proteome)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170413
[Lr] Data última revisão:
170413
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161222
[St] Status:MEDLINE
[do] DOI:10.1007/s11262-016-1416-9


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[PMID]:27453481
[Au] Autor:Matras M; Borzym E; Stone D; Way K; Stachnik M; Maj-Paluch J; Palusinska M; Reichert M
[Ad] Endereço:Department of Fish Diseases, National Veterinary Research Institute, 24-100, Pulawy, Poland.
[Ti] Título:Carp edema virus in Polish aquaculture - evidence of significant sequence divergence and a new lineage in common carp Cyprinus carpio (L.).
[So] Source:J Fish Dis;40(3):319-325, 2017 Mar.
[Is] ISSN:1365-2761
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Fish samples initially collected by local veterinarians on the common and koi carp farms in Poland between 2013 and 2015 as part of a KHV surveillance programme, when the water temperature was between 16 and 26 °C, and were also tested for CEV by qPCR. A partial 478 nucleotide fragment of the 4a gene was subsequently generated from 17 qPCR-positive common carp Cyprinus carpio samples from 36 farm sites tested during the period. Sequence alignments and analysis revealed the presence of CEV in Poland both in common carp as well as in koi carp farms, and phylogenetic analysis assigned the Polish CEV sequences into three distinct genogroups. A lineage which includes the original sequences obtained from koi carp in Japan (genogroup II) included sequences from both koi carp and common carp, and the second lineage (genogroup I) contained sequences from common carp only. A third lineage (genogroup III) which was more closely related to the genogroup II also consisted of sequences from common carp only. The latter represents a lineage of CEV not previously described in the literature.
[Mh] Termos MeSH primário: Carpas
Doenças dos Peixes/epidemiologia
Infecções por Poxviridae/veterinária
Poxviridae/fisiologia
[Mh] Termos MeSH secundário: Animais
Doenças dos Peixes/virologia
Filogenia
Polônia/epidemiologia
Reação em Cadeia da Polimerase/veterinária
Poxviridae/genética
Infecções por Poxviridae/epidemiologia
Infecções por Poxviridae/virologia
Análise de Sequência de DNA/veterinária
Proteínas Virais/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Viral Proteins)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170522
[Lr] Data última revisão:
170522
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160726
[St] Status:MEDLINE
[do] DOI:10.1111/jfd.12518


  10 / 1746 MEDLINE  
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[PMID]:27836975
[Au] Autor:Nuth M; Guan H; Ricciardi RP
[Ad] Endereço:From the Department of Microbiology, School of Dental Medicine and.
[Ti] Título:A Conserved Tripeptide Sequence at the C Terminus of the Poxvirus DNA Processivity Factor D4 Is Essential for Protein Integrity and Function.
[So] Source:J Biol Chem;291(53):27087-27097, 2016 12 30.
[Is] ISSN:1083-351X
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Vaccinia virus (VACV) is a poxvirus, and the VACV D4 protein serves both as a uracil-DNA glycosylase and as an essential component required for processive DNA synthesis. The VACV A20 protein has no known catalytic function itself but associates with D4 to form the D4-A20 heterodimer that functions as the poxvirus DNA processivity factor. The heterodimer enables the DNA polymerase to efficiently synthesize extended strands of DNA. Upon characterizing the interaction between D4 and A20, we observed that the C terminus of D4 is susceptible to perturbation. Further analysis demonstrated that a conserved hexapeptide stretch at the extreme C terminus of D4 is essential for maintaining protein integrity, as assessed by its requirement for the production of soluble recombinant protein that is functional in processive DNA synthesis. From the known crystal structures of D4, the C-terminal hexapeptide is shown to make intramolecular contact with residues spanning the inner core of the protein. Our mutational analysis revealed that a tripeptide motif ( GFI ) within the hexapeptide comprises apparent residues necessary for the contact. Prediction of protein disorder identified the hexapeptide and several regions upstream of Gly that comprise residues of the interface surfaces of the D4-A20 heterodimer. Our study suggests that GFI anchors these potentially dynamic upstream regions of the protein to maintain protein integrity. Unlike uracil-DNA glycosylases from diverse sources, where the C termini are disordered and do not form comparable intramolecular contacts, this feature may be unique to orthopoxviruses.
[Mh] Termos MeSH primário: DNA Viral/genética
Fragmentos de Peptídeos/química
Fragmentos de Peptídeos/metabolismo
Poxviridae/metabolismo
Proteínas Virais/química
Proteínas Virais/metabolismo
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Cristalografia por Raios X
Replicação do DNA
DNA Polimerase Dirigida por DNA/química
DNA Polimerase Dirigida por DNA/genética
DNA Polimerase Dirigida por DNA/metabolismo
Modelos Moleculares
Mutagênese Sítio-Dirigida
Mutação/genética
Fragmentos de Peptídeos/genética
Conformação Proteica
Domínios Proteicos
Homologia de Sequência de Aminoácidos
Uracila-DNA Glicosidase/química
Uracila-DNA Glicosidase/genética
Uracila-DNA Glicosidase/metabolismo
Proteínas Virais/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL
[Nm] Nome de substância:
0 (DNA, Viral); 0 (Peptide Fragments); 0 (Viral Proteins); EC 2.7.7.7 (DNA-Directed DNA Polymerase); EC 3.2.2.- (Uracil-DNA Glycosidase)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161113
[St] Status:MEDLINE
[do] DOI:10.1074/jbc.M116.761908



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