scholarly journals Efficient Transmission of Two Different Sheep Scrapie Isolates in Transgenic Mice Expressing the Ovine PrP Gene

2001 ◽  
Vol 75 (11) ◽  
pp. 5328-5334 ◽  
Author(s):  
Carole Crozet ◽  
Frederic Flamant ◽  
Anna Bencsik ◽  
Denise Aubert ◽  
Jacques Samarut ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Prion Protein ◽  
Neuron Specific Enolase ◽  
Open Reading Frame ◽  
Protein Accumulation ◽  
Spongiform Encephalopathies ◽  
Reading Frame ◽  
Neurological Signs ◽  
Prp Gene ◽  
Sheep Scrapie

ABSTRACT We produced transgenic mice expressing the sheep prion protein to obtain a sensitive model for sheep spongiform encephalopathies (scrapie). The complete open reading frame, with alanine, arginine, and glutamine at susceptibility codons 136, 154, and 171, respectively, was inserted downstream from the neuron-specific enolase promoter. A mouse line, Tg(OvPrP4), devoid of the murine PrP gene, was obtained by crossing with PrP knockout mice. Tg(OvPrP4) mice were shown to selectively express sheep PrP in their brains, as demonstrated in mRNA and protein analysis. We showed that these mice were susceptible to infection by sheep scrapie following intracerebral inoculation with two natural sheep scrapie isolates, as demonstrated not only by the occurrence of neurological signs but also by the presence of the spongiform changes and abnormal prion protein accumulation in their brains. Mean times to death of 238 and 290 days were observed with these isolates, but the clinical course of the disease was strikingly different in the two cases. One isolate led to a very early onset of neurological signs which could last for prolonged periods before death. Independently of the incubation periods, some of the mice inoculated with this isolate showed low or undetectable levels of PrPsc, as detected by both Western blotting and immunohistochemistry. The development of experimental scrapie in these mice following inoculation of the scrapie infectious agent further confirms that neuronal expression of the PrP open reading frame alone is sufficient to mediate susceptibility to spongiform encephalopathies. More importantly, these mice provide a new and promising tool for studying the infectious agents in sheep spongiform encephalopathies.

Comparative Analysis of the Prion Protein Open Reading Frame Nucleotide Sequences of Two Wild Ruminants, the Moufflon and Golden Takin

Intervirology ◽  
2001 ◽  
Vol 44 (6) ◽  
pp. 359-363 ◽  
Author(s):  
Sung Wook Seo ◽  
Kazuhiro Hara ◽  
Atsutaka Kubosaki ◽  
Yukiko Nasu ◽  
Takuya Nishimura ◽  
...  
Keyword(s):  
Comparative Analysis ◽  
Prion Protein ◽  
Nucleotide Sequences ◽  
Open Reading Frame ◽  
Reading Frame ◽  
Wild Ruminants ◽  
Golden Takin

scholarly journals UL26-Deficient Human Cytomegalovirus Produces Virions with Hypophosphorylated pp28 Tegument Protein That Is Unstable within Newly Infected Cells

2006 ◽  
Vol 80 (7) ◽  
pp. 3541-3548 ◽  
Author(s):  
Joshua Munger ◽  
Dong Yu ◽  
Thomas Shenk
Keyword(s):  
Human Cytomegalovirus ◽  
Deletion Mutant ◽  
Open Reading Frame ◽  
Early Gene ◽  
Immediate Early ◽  
Tegument Protein ◽  
Protein Accumulation ◽  
Reading Frame ◽  
Tegument Proteins ◽  
Infected Cells

ABSTRACT The human cytomegalovirus UL26 open reading frame encodes proteins of 21 and 27 kDa that result from the use of two different in-frame initiation codons. The UL26 protein is a constituent of the virion and thus is delivered to cells upon viral entry. We have characterized a mutant of human cytomegalovirus in which the UL26 open reading frame has been deleted. The UL26 deletion mutant has a profound growth defect, the magnitude of which is dependent on the multiplicity of infection. Two very early defects were discovered. First, even though they were present in normal amounts within mutant virions, the UL99-coded pp28 and UL83-coded pp65 tegument proteins were present in reduced amounts at the earliest times assayed within newly infected cells; second, there was a delay in immediate-early mRNA and protein accumulation. Further analysis revealed that although wild-type levels of the pp28 tegument protein were present in UL26 deletion mutant virions, the protein was hypophosphorylated. We conclude that the UL26 protein influences the normal phosphorylation of at least pp28 in virions and possibly additional tegument proteins. We propose that the hypophosphorylation of tegument proteins causes their destabilization within newly infected cells, perhaps disrupting the normal detegumentation process and leading to a delay in the onset of immediate-early gene expression.

scholarly journals Transmission and characterization of bovine spongiform encephalopathy sources in two ovine transgenic mouse lines (TgOvPrP4 and TgOvPrP59)

2006 ◽  
Vol 87 (12) ◽  
pp. 3763-3771 ◽  
Author(s):  
C. Cordier ◽  
A. Bencsik ◽  
S. Philippe ◽  
D. Bétemps ◽  
F. Ronzon ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Bovine Spongiform Encephalopathy ◽  
Transgenic Mouse ◽  
Neuron Specific Enolase ◽  
Infected Sheep ◽  
Transmissible Spongiform Encephalopathies ◽  
Spongiform Encephalopathy ◽  
Wild Type ◽  
Spongiform Encephalopathies ◽  
Mouse Lines

Transgenic mice expressing the prion protein (PrP) of species affected by transmissible spongiform encephalopathies (TSEs) have recently been produced to facilitate experimental transmission of these diseases by comparison with wild-type mice. However, whilst wild-type mice have largely been described for the discrimination of different TSE strains, including differentiation of agents involved in bovine spongiform encephalopathy (BSE) and scrapie, this has been only poorly described in transgenic mice. Here, two ovine transgenic mouse lines (TgOvPrP4 and TgOvPrP59), expressing the ovine PrP (A136 R154 Q171) under control of the neuron-specific enolase promoter, were studied; they were challenged with brainstem or spinal cord from experimentally BSE-infected sheep (AA136 RR154 QQ171 and AA136 RR154 RR171 genotypes) or brainstem from cattle BSE and natural sheep scrapie. The disease was transmitted successfully from all of these sources, with a mean of approximately 300 days survival following challenge with material from two ARQ-homozygous BSE-infected sheep in TgOvPrP4 mice, whereas the survival period in mice challenged with material from the ARR-homozygous BSE-infected sheep was 423 days on average. It was shown that, in the two ovine transgenic mouse lines, the Western blot characteristics of protease-resistant PrP (PrPres) were similar, whatever the BSE source, with a low apparent molecular mass of the unglycosylated glycoform, a poor labelling by P4 monoclonal antibody and high proportions of the diglycosylated form. With all BSE sources, but not with scrapie, florid plaques were observed in the brains of mice from both transgenic lines. These data reinforce the potential of this recently developed experimental model for the discrimination of BSE from scrapie agents.

NO MSPI POLYMORPHISM IN THE OPEN READING FRAME OF THE PRP GENE IN PATIENTS WITH FAMILIAL CREUTZFELD-JACOB DISEASE

1988 ◽  
Vol 2 (3) ◽  
pp. 311 ◽  
Author(s):  
D. GOLDGABER ◽  
J. W. TEENER ◽  
L. G. GOLDFARB ◽  
D. M. ASHER ◽  
P. W. BROWN ◽  
...  
Keyword(s):  
Open Reading Frame ◽  
Reading Frame ◽  
Mspi Polymorphism ◽  
Prp Gene

scholarly journals Prion protein genetics: Host PrP control of TSE susceptibility

2005 ◽  
Vol 27 (4) ◽  
pp. 20-23
Author(s):  
Wilfred Goldmann
Keyword(s):  
Prion Protein ◽  
Transmissible Spongiform Encephalopathies ◽  
Spongiform Encephalopathy ◽  
Spongiform Encephalopathies ◽  
Degenerative Disorders ◽  
Jakob Disease ◽  
The Central Nervous System ◽  
Important Host ◽  
Disease Family ◽  
Sheep Scrapie

TSEs (transmissible spongiform encephalopathies) are fatal, degenerative disorders of the central nervous system. The best-known members of this disease family are sheep scrapie, cattle BSE (bovine spongiform encephalopathy) and human CJD (Creutzfeldt–Jakob disease). By far the most important host gene in TSEs is the PrP (prion protein) gene. It modulates TSE susceptibility at many levels and is the crucial element in the treatment and eradication of these diseases. This article will highlight the advances in our understanding of PrP genetics in animals and man.

scholarly journals Release of Genotype 1 Hepatitis E Virus from Cultured Hepatoma and Polarized Intestinal Cells Depends on Open Reading Frame 3 Protein and Requires an Intact PXXP Motif

2010 ◽  
Vol 84 (18) ◽  
pp. 9059-9069 ◽  
Author(s):  
Suzanne U. Emerson ◽  
Hanh T. Nguyen ◽  
Udana Torian ◽  
Danielle Burke ◽  
Ronald Engle ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Cultured Cells ◽  
Hepatitis E ◽  
Open Reading Frame ◽  
Intestinal Cells ◽  
Protein Accumulation ◽  
Genotype 1 ◽  
Virus Genotype ◽  
Reading Frame ◽  
Orf3 Protein

ABSTRACT Hepatitis E virus genotype 1 strain Sar55 replicated in subcloned Caco-2 intestinal cells and Huh7 hepatoma cells that had been transfected with in vitro transcribed viral genomes, and hepatitis E virions were released into the culture medium of both cell lines. Virus egress from cells depended on open reading frame 3 (ORF3) protein, and a proline-rich sequence in ORF3 was important for egress from cultured cells and for infection of macaques. Both intracellular ORF3 protein accumulation and virus release occurred at the apical membrane of polarized Caco-2 cells. ORF3 protein and lipids were intimately associated with virus particles produced in either cell line; ORF2 epitopes were masked in these particles and could not be immunoprecipitated with anti-ORF2.

scholarly journals Absence of single nucleotide polymorphisms (SNPs) in the open reading frame (ORF) of the prion protein gene (PRNP) in a large sampling of various chicken breeds

BMC Genomics ◽  
2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Yong-Chan Kim ◽  
Sae-Young Won ◽  
Byung-Hoon Jeong
Keyword(s):  
Prion Protein ◽  
Prion Disease ◽  
Protein Gene ◽  
Open Reading Frame ◽  
High Dose ◽  
Prion Protein Gene ◽  
Genetic Characteristics ◽  
Reading Frame ◽  
Disease Resistant ◽  
Chicken Breeds

Abstract Background Prion diseases are zoonotic diseases with a broad infection spectrum among mammalian hosts and are caused by the misfolded prion protein (PrPSc) derived from the normal prion protein (PrPC), which encodes the prion protein gene (PRNP). Currently, although several prion disease-resistant animals have been reported, a high dose of prion agent inoculation triggers prion disease infection in these disease-resistant animals. However, in chickens, natural prion disease-infected cases have not been reported, and experimental challenges with prion agents have failed to cause infection. Unlike other prion disease-resistant animals, chickens have shown perfect resistance to prion disease thus far. Thus, investigation of the chicken PRNP gene could improve for understanding the mechanism of perfect prion-disease resistance. Here, we investigated the genetic characteristics of the open reading frame (ORF) of the chicken PRNP gene in a large sampling of various chicken breeds. Results We found only tandem repeat deletion polymorphisms of the chicken PRNP ORF in the 4 chicken breeds including 106 Dekalb White, 100 Ross, 98 Ogolgye and 100 Korean native chickens. In addition, the distribution of chicken insertion/deletion polymorphisms was significantly different among the 4 chicken breeds. Finally, we found significant differences in the number of PRNP SNPs between prion disease-susceptible species and prion disease-resistant species. Notably, chickens lack SNPs in the ORF of the prion protein. Conclusion In this study, we found that the absence of SNPs in the chicken PRNP ORF is a notable feature of animals with perfect resistant to prion disease.

scholarly journals Stability of BSE infectivity towards heat treatment even after proteolytic removal of prion protein

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Jan P. M. Langeveld ◽  
Anne Balkema-Buschmann ◽  
Dieter Becher ◽  
Achim Thomzig ◽  
Romolo Nonno ◽  
...  
Keyword(s):  
Heat Treatment ◽  
Prion Protein ◽  
Bovine Brain ◽  
Cellular Prion Protein ◽  
Transmissible Spongiform Encephalopathies ◽  
Spongiform Encephalopathies ◽  
Similar Treatment ◽  
Bank Voles ◽  
Before And After ◽  
Sheep Scrapie

AbstractThe unconventional infectious agents of transmissible spongiform encephalopathies (TSEs) are prions. Their infectivity co-appears with PrPSc, aberrant depositions of the host’s cellular prion protein (PrPC). Successive heat treatment in the presence of detergent and proteolysis by a keratinase from Bacillus licheniformis PWD-1 was shown before to destroy PrPSc from bovine TSE (BSE) and sheep scrapie diseased brain, however data regarding expected reduction of infectivity were still lacking. Therefore, transgenic Tgbov XV mice which are highly BSE susceptible were used to quantify infectivity before and after the bovine brain treatment procedure. Also four immunochemical analyses were applied to compare the levels of PrPSc. After heating at 115 °C with or without subsequent proteolysis, the original BSE infectivity of 106.2–6.4 ID50 g−1 was reduced to a remaining infectivity of 104.6–5.7 ID50 g−1 while strain characteristics were unaltered, even after precipitation with methanol. Surprisingly, PrPSc depletion was 5–800 times higher than the loss of infectivity. Similar treatment was applied on other prion strains, which were CWD1 in bank voles, 263 K scrapie in hamsters and sheep PG127 scrapie in tg338 ovinized mice. In these strains however, infectivity was already destroyed by heat only. These findings show the unusual heat resistance of BSE and support a role for an additional factor in prion formation as suggested elsewhere when producing prions from PrPC. Leftover material in the remaining PrPSc depleted BSE preparation offers a unique substrate for searching additional elements for prion infectivity and improving our concept about the nature of prions.

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