scholarly journals M Segment-Based Minigenomes and Virus-Like Particle Assays as an Approach To Assess the Potential of Tick-BornePhlebovirusGenome Reassortment

2018 ◽  
Vol 93 (6) ◽  
Author(s):  
Veronica V. Rezelj ◽  
Timothy J. Mottram ◽  
Joseph Hughes ◽  
Richard M. Elliott ◽  
Alain Kohl ◽  
...  
Keyword(s):  
Genome Segment ◽  
Emerging Pathogens ◽  
Virus Like Particle ◽  
A Genome ◽  
Large Expansion ◽  
Heterologous Virus ◽  
Theory Result ◽  
L Proteins ◽  
Genome Reassortment ◽  
Reassortant Viruses

ABSTRACTBunyaviruses have a tripartite negative-sense RNA genome. Due to the segmented nature of these viruses, if two closely related viruses coinfect the same host or vector cell, it is possible that RNA segments from either of the two parental viruses will be incorporated into progeny virions to give reassortant viruses. Little is known about the ability of tick-borne phleboviruses to reassort. The present study describes the development of minigenome assays for the tick-borne viruses Uukuniemi phlebovirus (UUKV) and Heartland phlebovirus (HRTV). We used these minigenome assays in conjunction with the existing minigenome system of severe fever with thrombocytopenia syndrome (SFTS) phlebovirus (SFTSV) to assess the abilities of viral N and L proteins to recognize, transcribe, and replicate the M segment-based minigenome of a heterologous virus. The highest minigenome activity was detected with the M segment-based minigenomes of cognate viruses. However, our findings indicate that several combinations utilizing N and L proteins of heterologous viruses resulted in M segment minigenome activity. This suggests that the M segment untranslated regions (UTRs) are recognized as functional promoters of transcription and replication by the N and L proteins of related viruses. Further, virus-like particle assays demonstrated that HRTV glycoproteins can package UUKV and SFTSV S and L segment-based minigenomes. Taken together, these results suggest that coinfection with these viruses could lead to the generation of viable reassortant progeny. Thus, the tools developed in this study could aid in understanding the role of genome reassortment in the evolution of these emerging pathogens in an experimental setting.IMPORTANCEIn recent years, there has been a large expansion in the number of emerging tick-borne viruses that are assigned to thePhlebovirusgenus. Bunyaviruses have a tripartite segmented genome, and infection of the same host cell by two closely related bunyaviruses can, in theory, result in eight progeny viruses with different genome segment combinations. We used genome analogues expressing reporter genes to assess the abilities ofPhlebovirusnucleocapsid protein and RNA-dependent RNA polymerase to recognize the untranslated region of a genome segment of a related phlebovirus, and we used virus-like particle assays to assess whether viral glycoproteins can package genome analogues of related phleboviruses. Our results provide strong evidence that these emerging pathogens could reassort their genomes if they were to meet in nature in an infected host or vector. This reassortment process could result in viruses with new pathogenic properties.

scholarly journals A Lymphatic Mechanism of Rotavirus Extraintestinal Spread in the Neonatal Mouse

2003 ◽  
Vol 77 (22) ◽  
pp. 12352-12356 ◽  
Author(s):  
Eric C. Mossel ◽  
Robert F. Ramig
Keyword(s):  
Genome Segment ◽  
Neonatal Mouse ◽  
Mesenteric Lymph Nodes ◽  
Reassortant Virus ◽  
Peripheral Tissues ◽  
Mesenteric Lymph ◽  
Oral Inoculation ◽  
Reassortant Viruses ◽  
Lymphatic Pathway ◽  
Virus Strains

ABSTRACT We used the neonatal mouse model of rotavirus infection and virus strains SA11-clone 4 (SA11-Cl4) and Rhesus rotavirus (RRV) to examine the mechanism of the extraintestinal spread of viruses following oral inoculation. The spread-competent viruses, RRV and reassortant R7, demonstrated a temporal progression from the intestine, to the terminal ileum, to the mesenteric lymph nodes (MLN), and to the peripheral tissues. SA11-Cl4 was not found outside the intestine. Reassortant virus S7, which was unable to reach the liver in previous studies (E. C. Mossel and R. F. Ramig, J. Virol. 76:6502-6509, 2002), was recovered from 60% of the MLN, suggesting that there are multiple determinants for the spread of virus from the intestine to the MLN. Phenotypic segregation analysis identified RRV genome segment 6 (VP6) as a secondary determinant of the spread of virus to the MLN (P = 0.02) in reassortant viruses containing segment 7 from the spread-incompetent parent. These data suggest that in the orally infected neonatal mouse, the extraintestinal spread of rotavirus occurs via a lymphatic pathway, and the spread phenotype is primarily determined by NSP3 and can be modified by VP6.

scholarly journals Generation of simian rotavirus reassortants with diverse VP4 genes using reverse genetics

2019 ◽  
Vol 100 (12) ◽  
pp. 1595-1604 ◽  
Author(s):  
Alexander Falkenhagen ◽  
Corinna Patzina-Mehling ◽  
Antje Rückner ◽  
Thomas W. Vahlenkamp ◽  
Reimar Johne
Keyword(s):  
Viral Entry ◽  
Reverse Genetics ◽  
Genome Segment ◽  
Cell Tropism ◽  
Outer Capsid Protein ◽  
High Genetic Diversity ◽  
Cytopathic Effects ◽  
Reassortant Viruses ◽  
Outer Capsid ◽  
Virus Strains

Species A rotaviruses (RVAs) are a major cause of gastroenteritis in animals and humans. Their genome consists of 11 segments of dsRNA, and reassortment events between animal and human strains can contribute to the high genetic diversity of RVAs. We used a plasmid-based reverse genetics system to investigate the reassortment potential of the genome segment encoding the viral outer capsid protein VP4, which is a major antigenic determinant, mediates viral entry and plays an important role in host cell tropism. We rescued reassortant viruses containing VP4 from porcine, bovine, bat, pheasant or chicken RVA strains in the backbone of simian strain SA11. The VP4 reassortants could be stably passaged in MA-104 cells and induced cytopathic effects. However, analysis of growth kinetics revealed marked differences in replication efficiency. Our results show that the VP4-encoding genome segment has a high reassortment potential, even between virus strains from highly divergent species. This can result in replication-competent reassortants with new genomic, growth and antigenic features.

scholarly journals An Enhanced Heterologous Virus-Like Particle for Human Papillomavirus Type 16 Tumour Immunotherapy

PLoS ONE ◽  
2013 ◽  
Vol 8 (6) ◽  
pp. e66866 ◽  
Author(s):  
Khairunadwa Jemon ◽  
Vivienne Young ◽  
Michelle Wilson ◽  
Sara McKee ◽  
Vernon Ward ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Human Papillomavirus Type 16 ◽  
Tumour Immunotherapy ◽  
Virus Like Particle ◽  
Heterologous Virus

scholarly journals Ribosome Profiling Uncovers the Role of uORFs in Translational Control of Gene Expression during Erythroblast Differentiation

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 2658-2658
Author(s):  
Klaske A.M.H. Thiadens ◽  
Eleonora de Klerk ◽  
Ivo F.A.C. Fokkema ◽  
Peter A.C. ‘t Hoen ◽  
Marieke von Lindern
Keyword(s):  
Translation Initiation ◽  
Translational Control ◽  
Mrna Translation ◽  
Ribosome Profiling ◽  
Start Codon ◽  
Translation Efficiency ◽  
Control Of Gene Expression ◽  
Genome Wide ◽  
A Genome ◽  
Large Expansion

Abstract The erythroid progenitor compartment possesses a large expansion capacity, both in vivo and in vitro, which enables a rapid restoration of peripheral erythrocytes following severe blood loss. This expansion is tightly regulated to maintain erythrocyte numbers between narrow boundaries, and to balance expansion of the erythroid compartment against the availability of iron for heme and haemoglobin production. We previously observed that control of mRNA translation is crucial for expansion of the erythroid compartment. We also showed that translation of specific transcripts is impaired in Diamond Blackfan Anemia (DBA), a severe congenital anemia due to defective ribosome biosynthesis. Transcripts can be subject to translational control through domains in the 5’- or 3’UTR, including secondary structures, protein binding sequences and upstream open reading frames (uORFs). The presence of uORFs, including those starting at non-AUG codons in the 5’UTR, may alter the level of mRNA translation, but may also result in the expression of alternative protein isoforms because translation initiation may be redirected to more downstream start codons. The aim of our current studies is to provide a genome wide map of mRNA translation efficiency during erythropoiesis that can be used to investigate defective mRNA translation in, for instance, DBA. Ribosome profiling is a genome wide high-throughput sequencing technology for global mapping of translation initiation sites that allows translation analysis with codon resolution at the genome wide level. We first investigated translational changes occurring during differentiation of mouse erythroblasts. We used p53-deficient, growth factor dependent and differentiation competent immortalized erythroblast cultures that were expanded in presence of erythropoietin (Epo), stem cell factor (SCF) and glucocorticoids as T0, and subsequently differentiated the cells in presence of Epo for 17 and 46 hours (T17, and T46 samples). To obtain ribosome footprints, the cells were treated for 7 minutes with harringtonin or solvent, and subsequently for 5 minutes with cycloheximide, which arrests translation by stabilizing the ribosomes at translation initiation codons, or on all codons, respectively. We used optimized protocols for ribosome footprinting and data analysis, and focused the analysis on transcripts containing uORFs. First we performed a qualitative analysis of start codon usage. The ribosome footprint data proved to be superior to previously used polyribosome recruitment. In some cases polysome recruitment appeared to represent translation of an uORFs while the protein coding ORF is hardly translated (e.g. Csf2rb2, Puma). In another set of transcripts, we found uORFs that are differentially translated during differentiation, and thereby regulate differential translation from a downstream start codon (e.g. Klf3, Use1, CD47, Kell). Finally, comparison of ribosome footprints determined in erythroblasts and in myoblasts/myotubes revealed tissue specific translation regulation of otherwise ubiquitously expressed transcripts among which transcripts encoding ribosomal proteins. Second, we will perform quantitative analysis of mRNA translation in erythropoiesis through the comparison of ribosome footprint reads in an ORF with total mRNA reads obtained from total mRNA sequencing of the same sample. The obtained insight in transcript specific translation at codon resolution is of great value to understand many cellular processes during erythropoiesis, and will be of particular interest to understand responses to iron availability and reactive oxygen species that particularly affect translation of transcripts harboring uORFs. Disclosures No relevant conflicts of interest to declare.

scholarly journals Characterization of Donor Genome Segments of BC2 and BC4 Way Rarem x Oryzica Llanos-5 Progenies Detected by SNP Markers

2016 ◽  
Vol 8 (1) ◽  
pp. 1
Author(s):  
Wening Enggarini ◽  
Surjono H. Sudjahjo ◽  
Trikoesoemaningtyas Trikoesoemaningtyas ◽  
Sriani Sujiprihati ◽  
Utut Widyastuti ◽  
...  
Keyword(s):  
Rice Genome ◽  
Snp Markers ◽  
Genome Segment ◽  
Recurrent Parent ◽  
Donor Genome ◽  
Simple Method ◽  
Advanced Backcross ◽  
Donor Segment ◽  
A Genome ◽  
Donor Parent

<p>Plant breeders<br />make a succession of backcrosses to introgress a character<br />from a donor parent into genomic background of a recurrent<br />parent. In several backcrossing, the proportion of a genome<br />tends to return almost fully to recurrent parent, except the<br />small donor genome segment harboring the character of<br />interest. The estimation of the proportion donor segment<br />through backcross generations has been analyzed<br />theoretically using complex mathematical simulations. In<br />this study, the proportion of donor introgression segments<br />were directly analyzed in advanced backcross populations,<br />BC2F7 and BC4F2. The analysis was done by using a set of<br />single nucleotide polymorphism (SNP) markers covering the<br />entire rice genome. Of the 384 SNP markers we found 124<br />markers which provide polymorphism between recurrent<br />parent, Way Rarem and Oryzica Llanos-5 as donor parent.<br />But only 55 SNP markers could detect Oryzica Llanos-5<br />alleles in BC2F7 and BC4F2 progenies. The result of this<br />analysis demonstrated that the average of donor segment<br />number was 14.5 in BC2F7 and 12.3 in BC4F2. It was reduced<br />15% from BC2F7 to BC4F2. The average of donor segment<br />length was 31.2 cM (centiMorgan) in BC2F7 and 8.79 cM in<br />BC4F2. It was decreased 72% during twice backcrossing. The<br />average of donor genome size was 343.95 cM in BC2F7 and<br />71.35 cM in BC4F2, which means there was 79% decrease<br />from BC2F7 to BC4F2. These results offered a simple method<br />to describe the proportion of target genome segment from<br />donor parent. It was required as one of the main selection<br />criteria in backcross programs.</p>

H5N1 influenza virus-like particle vaccine protects mice from heterologous virus challenge better than whole inactivated virus

2015 ◽  
Vol 200 ◽  
pp. 9-18 ◽  
Author(s):  
Zhiguang Ren ◽  
Xianliang Ji ◽  
Lingnan Meng ◽  
Yurong Wei ◽  
Tiecheng Wang ◽  
...  
Keyword(s):  
Influenza Virus ◽  
H5n1 Influenza Virus ◽  
Virus Challenge ◽  
H5n1 Influenza ◽  
Virus Like Particle ◽  
Heterologous Virus ◽  
Better Than

Immune Electron Microscopy (IEM) of a Canine Adeno-Associated Virus

1974 ◽  
Vol 32 ◽  
pp. 256-257
Author(s):  
Gunter F. Thomas ◽  
M. David Hoggan
Keyword(s):  
Electron Microscopy ◽  
Cell Cultures ◽  
Complement Fixation ◽  
Hepatitis Virus ◽  
Wide Distribution ◽  
Immune Electron Microscopy ◽  
Adeno Associated Virus ◽  
Virus Like Particle ◽  
Dog Kidney ◽  
Green Monkeys

In 1968, Sugimura and Yanagawa described a small 25 nm virus like particle in association with the Matsuda strain of infectious canine hepatitis virus (ICHV). Domoto and Yanagawa showed that this particle was dependent on ICHV for its replication in primary dog kidney cell cultures (PDK) and was resistant to heating at 70°C for 10 min, and concluded that it was a canine adeno-associated virus (CAAV). Later studies by Onuma and Yanagawa compared CAAV with the known human serotypes (AAV 1, 2, 3) and AAV-4, known to be associated with African Green Monkeys. Using the complement fixation (CF) test, they found that CAAV was serologically related to AAV-3 and had wide distribution in the dog population of Japan.

Hepatitis A Antigen in Liver, Bile and Stool

1975 ◽  
Vol 33 ◽  
pp. 340-341
Author(s):  
D.R. Jackson ◽  
J.H. Hoofnagle ◽  
A.N. Schulman ◽  
J.L. Dienstag ◽  
R.H. Purcell ◽  
...  
Keyword(s):  
Hepatitis A ◽  
Liver Enzymes ◽  
Hepatitis A Virus ◽  
Type A ◽  
Initial Study ◽  
Virus Like Particle ◽  
Elevated Liver Enzymes ◽  
Ag Particles ◽  
Ag Particle ◽  
Human Stool

Using immune electron microscopy Feinstone et. al. demonstrated the presence of a 27 nm virus-like particle in acute-phase stools of patients with viral hepatitis, type A, These hepatitis A antigen (HA Ag) particles were aggregated by convalescent serum from patients with type A hepatitis but not by pre-infection serum. Subsequently Dienstag et. al. and Maynard et. al. produced acute hepatitis in chimpanzees by inoculation with human stool containing HA Ag. During the early acute disease, virus like particles antigenically, morphologically and biophysically identical to the human HA Ag particle were found in chimpanzee stool. Recently Hilleman et. al. have described similar particles in liver and serum of marmosets infected with hepatitis A virus (HAV). We have investigated liver, bile and stool from chimpanzees and marmosets experimentally infected with HAV. In an initial study, a chimpanzee (no.785) inoculated with HA Ag-containing stool developed elevated liver enzymes 21 days after exposure.

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