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1.
Biomolecules ; 13(12)2023 12 13.
Article in English | MEDLINE | ID: mdl-38136658

ABSTRACT

Human neurodegenerative diseases associated with the misfolding of the alpha-synuclein (aS) protein (synucleinopathies) are similar to prion diseases to the extent that lesions are spread by similar molecular mechanisms. In a transgenic mouse model (M83) overexpressing a mutated (A53T) form of human aS, we had previously found that Protein Misfolding Cyclic Amplification (PMCA) triggered the aggregation of aS, which is associated with a high resistance to the proteinase K (PK) digestion of both human and murine aS, a major hallmark of the disease-associated prion protein. In addition, PMCA was also able to trigger the aggregation of murine aS in C57Bl/6 mouse brains after seeding with sick M83 mouse brains. Here, we show that intracerebral inoculations of M83 mice with C57Bl/6-PMCA samples strikingly shortens the incubation period before the typical paralysis that develops in this transgenic model, demonstrating the pathogenicity of PMCA-aggregated murine aS. In the hind brain regions of these sick M83 mice containing lesions with an accumulation of aS phosphorylated at serine 129, aS also showed a high PK resistance in the N-terminal part of the protein. In contrast to M83 mice, old APPxM83 mice co-expressing human mutated amyloid precursor and presenilin 1 proteins were seen to have an aggregation of aS, especially in the cerebral cortex, hippocampus and striatum, which also contained the highest load of aS phosphorylated at serine 129. This was proven by three techniques: a Western blot analysis of PK-resistant aS; an ELISA detection of aS aggregates; or the identification of aggregates of aS using immunohistochemical analyses of cytoplasmic/neuritic aS deposits. The results obtained with the D37A6 antibody suggest a higher involvement of murine aS in APPxM83 mice than in M83 mice. Our study used novel tools for the molecular study of synucleinopathies, which highlight similarities with the molecular mechanisms involved in prion diseases.


Subject(s)
Prion Diseases , Synucleinopathies , Animals , Humans , Mice , alpha-Synuclein/genetics , alpha-Synuclein/metabolism , Brain/metabolism , Mice, Transgenic , Peptide Hydrolases/metabolism , Prion Diseases/pathology , Serine/metabolism , Synucleinopathies/metabolism
2.
Sci Adv ; 9(37): eadi7838, 2023 09 15.
Article in English | MEDLINE | ID: mdl-37703372

ABSTRACT

Tubulin posttranslational modifications represent an important mechanism involved in the regulation of microtubule functions. The most widespread among them are detyrosination, α∆2-tubulin, and polyglutamylation. Here, we describe a family of tubulin-modifying enzymes composed of two closely related proteins, KIAA0895L and KIAA0895, which have tubulin metallocarboxypeptidase activity and thus were termed TMCP1 and TMCP2, respectively. We show that TMCP1 (also known as MATCAP) acts as α-tubulin detyrosinase that also catalyzes α∆2-tubulin. In contrast, TMCP2 preferentially modifies ßI-tubulin by removing three amino acids from its C terminus, generating previously unknown ßI∆3 modification. We show that ßI∆3-tubulin is mostly found on centrioles and mitotic spindles and in cilia. Moreover, we demonstrate that TMCPs also remove posttranslational polyglutamylation and thus act as tubulin deglutamylases. Together, our study describes the identification and comprehensive biochemical analysis of a previously unknown type of tubulin-modifying enzymes involved in the processing of α- and ß-tubulin C-terminal tails and deglutamylation.


Subject(s)
Carboxypeptidases , Tubulin , Microtubules , Amino Acids , Centrioles
3.
Sci Rep ; 11(1): 4058, 2021 02 18.
Article in English | MEDLINE | ID: mdl-33603091

ABSTRACT

Unlike variant Creutzfeldt-Jakob disease prions, sporadic Creutzfeldt-Jakob disease prions have been shown to be difficult to amplify in vitro by protein misfolding cyclic amplification (PMCA). We assessed PMCA of pathological prion protein (PrPTSE) from 14 human sCJD brain samples in 3 substrates: 2 from transgenic mice expressing human prion protein (PrP) with either methionine (M) or valine (V) at position 129, and 1 from bank voles. Brain extracts representing the 5 major clinicopathological sCJD subtypes (MM1/MV1, MM2, MV2, VV1, and VV2) all triggered seeded PrPTSE amplification during serial PMCA with strong seed- and substrate-dependence. Remarkably, bank vole PrP substrate allowed the propagation of all sCJD subtypes with preservation of the initial molecular PrPTSE type. In contrast, PMCA in human PrP substrates was accompanied by a PrPTSE molecular shift during heterologous (M/V129) PMCA reactions, with increased permissiveness of V129 PrP substrate to in vitro sCJD prion amplification compared to M129 PrP substrate. Combining PMCA amplification sensitivities with PrPTSE electrophoretic profiles obtained in the different substrates confirmed the classification of 4 distinct major sCJD prion strains (M1, M2, V1, and V2). Finally, the level of sensitivity required to detect VV2 sCJD prions in cerebrospinal fluid was achieved.


Subject(s)
Creutzfeldt-Jakob Syndrome/metabolism , Prions/metabolism , Animals , Arvicolinae/metabolism , Humans , Mice , Mice, Transgenic , Prion Proteins/metabolism , Protein Folding , Proteostasis Deficiencies/metabolism
4.
Nat Commun ; 11(1): 2818, 2020 06 04.
Article in English | MEDLINE | ID: mdl-32499524

ABSTRACT

In eukaryotes, trimethylation of lysine 9 on histone H3 (H3K9) is associated with transcriptional silencing of transposable elements (TEs). In drosophila ovaries, this heterochromatic repressive mark is thought to be deposited by SetDB1 on TE genomic loci after the initial recognition of nascent transcripts by PIWI-interacting RNAs (piRNAs) loaded on the Piwi protein. Here, we show that the nucleosome remodeler Mi-2, in complex with its partner MEP-1, forms a subunit that is transiently associated, in a MEP-1 C-terminus-dependent manner, with known Piwi interactors, including a recently reported SUMO ligase, Su(var)2-10. Together with the histone deacetylase Rpd3, this module is involved in the piRNA-dependent TE silencing, correlated with H3K9 deacetylation and trimethylation. Therefore, drosophila piRNA-mediated transcriptional silencing involves three epigenetic effectors, a remodeler, Mi-2, an eraser, Rpd3 and a writer, SetDB1, in addition to the Su(var)2-10 SUMO ligase.


Subject(s)
Adenosine Triphosphatases/metabolism , Autoantigens/metabolism , Drosophila Proteins/metabolism , Heterochromatin/chemistry , Histone Deacetylase 1/metabolism , Nucleosomes/metabolism , RNA, Small Interfering/metabolism , Animals , Argonaute Proteins/metabolism , Drosophila melanogaster , Epigenesis, Genetic , Female , Gene Expression Regulation , Gene Silencing , Histones/chemistry , Ovary/metabolism , Protein Inhibitors of Activated STAT
5.
mSphere ; 5(1)2020 Jan 29.
Article in English | MEDLINE | ID: mdl-31996421

ABSTRACT

To date, approximately 500 iatrogenic Creutzfeldt-Jakob disease cases have been reported worldwide, most of them resulting from cadaveric dura mater graft and from the administration of prion-contaminated human growth hormone. The unusual resistance of prions to decontamination processes, their large tissue distribution, and the uncertainty about the prevalence of variant Creutzfeldt-Jakob disease (vCJD) in the general population lead to specific recommendations regarding identification of tissue at risk and reprocessing of reusable medical devices, including the use of dedicated treatment for prion inactivation. We previously described an in vitro assay, called Surf-PMCA, which allowed us to classify prion decontamination treatments according to their efficacy on vCJD prions by monitoring residual seeding activity (RSA). Here, we used a transgenic mouse line permissive to vCJD prions to study the correlation between the RSA measured in vitro and the in vivo infectivity. Implantation in mouse brains of prion-contaminated steel wires subjected to different decontamination procedures allows us to demonstrate a good concordance between RSA measured by Surf-PMCA (in vitro) and residual infectivity (in vivo). These experiments emphasize the strength of the Surf-PMCA method as a rapid and sensitive assay for the evaluation of prion decontamination procedures and also confirm the lack of efficacy of several marketed reagents on vCJD prion decontamination.IMPORTANCE Creutzfeldt-Jakob diseases are neurodegenerative disorders for which transmission linked to medical procedures have been reported in hundreds of patients. As prion diseases, they are characterized by an unusual resistance to conventional decontamination processes. Moreover, their large tissue distribution and the ability of prions to attach to many surfaces raised the risk of transmission in health care facilities. It is therefore of major importance that decontamination procedures applied to medical devices before their reprocessing are thoroughly validated for prion inactivation. We previously described an in vitro assay, which allowed us to classify accurately prion decontamination treatments according to their efficacy on variant Creutzfeldt-Jakob disease. The significance of this study is in demonstrating the concordance between previous in vitro results and infectivity studies in transgenic mice. Furthermore, commercial reagents currently used in hospitals were tested by both protocols, and we observed that most of them were ineffective on human prions.


Subject(s)
Creutzfeldt-Jakob Syndrome/pathology , Decontamination/methods , Equipment Contamination , Prion Proteins/chemistry , Animals , Creutzfeldt-Jakob Syndrome/etiology , Female , Humans , Iatrogenic Disease , Mice , Mice, Transgenic , Proteostasis Deficiencies/pathology
6.
FASEB J ; 33(11): 12073-12086, 2019 11.
Article in English | MEDLINE | ID: mdl-31370680

ABSTRACT

α-Synuclein (α-syn) protein aggregation is associated with several neurodegenerative disorders collectively referred to as synucleinopathies, including Parkinson's disease. We used protein misfolding cyclic amplification (PMCA) to study α-syn aggregation in brain homogenates of wild-type or transgenic mice expressing normal (D line) or A53T mutant (M83 line) human α-syn. We found that sonication-incubation cycles of M83 mouse brain gradually produce large quantities of SDS-resistant α-syn aggregates, involving both human and mouse proteins. These PMCA products, containing partially proteinase K-resistant α-syn species, are competent to accelerate the onset of neurologic symptoms after intracerebral inoculation to young M83 mice and to seed aggregate formation of α-syn following PMCA, including in D and wild-type mouse brain substrates. PMCA seeding activity in the M83 diseased brain correlates positively with regions mostly targeted by the α-syn pathology in this model. Our data indicate that similar to prions, PMCA can reproduce some characteristics of α-syn aggregation and seeded propagation in vitro in a complex milieu. This opens new opportunities for the molecular study of synucleinopathies.-Nicot, S., Verchère, J., Bélondrade, M., Mayran, C., Bétemps, D., Bougard, D., Baron, T. Seeded propagation of α-synuclein aggregation in mouse brain using protein misfolding cyclic amplification.


Subject(s)
Brain/metabolism , Gene Amplification , Protein Aggregates , Protein Aggregation, Pathological/genetics , Proteostasis Deficiencies/genetics , alpha-Synuclein/genetics , Animals , Brain/pathology , Humans , Mice, Inbred C57BL , Mice, Knockout , Mice, Transgenic , Mutation , Protein Aggregation, Pathological/metabolism , Proteostasis Deficiencies/metabolism , alpha-Synuclein/chemistry , alpha-Synuclein/metabolism
7.
Sci Transl Med ; 8(370): 370ra182, 2016 12 21.
Article in English | MEDLINE | ID: mdl-28003547

ABSTRACT

Variant Creutzfeldt-Jakob disease (vCJD) is a human prion disease resulting from the consumption of meat products contaminated by the agent causing bovine spongiform encephalopathy. Evidence supporting the presence of a population of silent carriers that can potentially transmit the disease through blood transfusion is increasing. The development of a blood-screening assay for both symptomatic vCJD patients and asymptomatic carriers is urgently required. We show that a diagnostic assay combining plasminogen-bead capture and protein misfolding cyclic amplification (PMCA) technologies consistently detected minute amounts of abnormal prion protein from French and British vCJD cases in the required femtomolar range. This assay allowed the blinded identification of 18 patients with clinical vCJD among 256 plasma samples from the two most affected countries, with 100% sensitivity [95% confidence interval (CI), 81.5 to 100%], 99.2% analytical specificity (95% CI, 95.9 to 100%), and 100% diagnostic specificity (95% CI, 96.5 to 100%). This assay also allowed the detection of silent carriage of prions 1.3 and 2.6 years before the clinical onset in two blood donors who later developed vCJD. These data provide a key step toward the validation of this PMCA technology as a blood-based diagnostic test for vCJD and support its potential for detecting presymptomatic patients, a prerequisite for limiting the risk of vCJD transmission through blood transfusion.


Subject(s)
Creutzfeldt-Jakob Syndrome/blood , Creutzfeldt-Jakob Syndrome/diagnosis , Hematologic Tests/methods , Prion Proteins/blood , France , Humans , Reproducibility of Results , Sensitivity and Specificity , Treatment Outcome , United Kingdom
8.
PLoS One ; 11(1): e0146833, 2016.
Article in English | MEDLINE | ID: mdl-26800081

ABSTRACT

The prevalence of variant Creutzfeldt-Jakob disease (vCJD) in the population remains uncertain, although it has been estimated that 1 in 2000 people in the United Kingdom are positive for abnormal prion protein (PrPTSE) by a recent survey of archived appendix tissues. The prominent lymphotropism of vCJD prions raises the possibility that some surgical procedures may be at risk of iatrogenic vCJD transmission in healthcare facilities. It is therefore vital that decontamination procedures applied to medical devices before their reprocessing are thoroughly validated. A current limitation is the lack of a rapid model permissive to human prions. Here, we developed a prion detection assay based on protein misfolding cyclic amplification (PMCA) technology combined with stainless-steel wire surfaces as carriers of prions (Surf-PMCA). This assay allowed the specific detection of minute quantities (10-8 brain dilution) of either human vCJD or ovine scrapie PrPTSE adsorbed onto a single steel wire, within a two week timeframe. Using Surf-PMCA we evaluated the performance of several reference and commercially available prion-specific decontamination procedures. Surprisingly, we found the efficiency of several marketed reagents to remove human vCJD PrPTSE was lower than expected. Overall, our results demonstrate that Surf-PMCA can be used as a rapid and ultrasensitive assay for the detection of human vCJD PrPTSE adsorbed onto a metallic surface, therefore facilitating the development and validation of decontamination procedures against human prions.


Subject(s)
Biological Assay/methods , Creutzfeldt-Jakob Syndrome/prevention & control , Cross Infection/prevention & control , Decontamination/methods , Prions/metabolism , Animals , Brain/pathology , Creutzfeldt-Jakob Syndrome/pathology , Creutzfeldt-Jakob Syndrome/transmission , Cross Infection/transmission , Equipment and Supplies , Humans , Limit of Detection , Mice , PrPSc Proteins/metabolism , Protein Folding , Scrapie/metabolism , Stainless Steel , Surface Properties , United Kingdom
9.
PLoS Pathog ; 11(8): e1005073, 2015 Aug.
Article in English | MEDLINE | ID: mdl-26241960

ABSTRACT

In prion diseases, synapse dysfunction, axon retraction and loss of neuronal polarity precede neuronal death. The mechanisms driving such polarization defects, however, remain unclear. Here, we examined the contribution of RhoA-associated coiled-coil containing kinases (ROCK), key players in neuritogenesis, to prion diseases. We found that overactivation of ROCK signaling occurred in neuronal stem cells infected by pathogenic prions (PrPSc) and impaired the sprouting of neurites. In reconstructed networks of mature neurons, PrPSc-induced ROCK overactivation provoked synapse disconnection and dendrite/axon degeneration. This overactivation of ROCK also disturbed overall neurotransmitter-associated functions. Importantly, we demonstrated that beyond its impact on neuronal polarity ROCK overactivity favored the production of PrPSc through a ROCK-dependent control of 3-phosphoinositide-dependent kinase 1 (PDK1) activity. In non-infectious conditions, ROCK and PDK1 associated within a complex and ROCK phosphorylated PDK1, conferring basal activity to PDK1. In prion-infected neurons, exacerbated ROCK activity increased the pool of PDK1 molecules physically interacting with and phosphorylated by ROCK. ROCK-induced PDK1 overstimulation then canceled the neuroprotective α-cleavage of normal cellular prion protein PrPC by TACE α-secretase, which physiologically precludes PrPSc production. In prion-infected cells, inhibition of ROCK rescued neurite sprouting, preserved neuronal architecture, restored neuronal functions and reduced the amount of PrPSc. In mice challenged with prions, inhibition of ROCK also lowered brain PrPSc accumulation, reduced motor impairment and extended survival. We conclude that ROCK overactivation exerts a double detrimental effect in prion diseases by altering neuronal polarity and triggering PrPSc accumulation. Eventually ROCK emerges as therapeutic target to combat prion diseases.


Subject(s)
PrPSc Proteins/metabolism , Prion Diseases/metabolism , Prion Diseases/pathology , rho-Associated Kinases/metabolism , Animals , Blotting, Western , Disease Models, Animal , Fluorescent Antibody Technique , Immunoprecipitation , Lab-On-A-Chip Devices , Mice , Mice, Inbred C57BL , Neurites/metabolism , Neurogenesis , PrPC Proteins/metabolism
10.
J Infect Dis ; 209(6): 950-9, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24218507

ABSTRACT

BACKGROUND: Sheep with prion protein (PrP) gene polymorphisms QQ171 and RQ171 were shown to be susceptible to the prion causing L-type bovine spongiform encephalopathy (L-BSE), although RQ171 sheep specifically propagated a distinctive prion molecular phenotype in their brains, characterized by a high molecular mass protease-resistant PrP fragment (HMM PrPres), distinct from L-BSE in QQ171 sheep. METHODS: The resulting infectious and biological properties of QQ171 and RQ171 ovine L-BSE prions were investigated in transgenic mice expressing either bovine or ovine PrP. RESULTS: In both mouse lines, ovine L-BSE transmitted similarly to cattle-derived L-BSE, with respect to survival periods, histopathology, and biochemical features of PrPres in the brain, as well as splenotropism, clearly differing from ovine classic BSE or from scrapie strain CH1641. Nevertheless and unexpectedly, HMM PrPres was found in the spleen of ovine PrP transgenic mice infected with L-BSE from RQ171 sheep at first passage, reminiscent, in lymphoid tissues only, of the distinct PrPres features found in RQ171 sheep brains. CONCLUSIONS: The L-BSE agent differs from both ovine classic BSE or CH1641 scrapie maintaining its specific strain properties after passage in sheep, although striking PrPres molecular changes could be found in RQ171 sheep and in the spleen of ovine PrP transgenic mice.


Subject(s)
Encephalopathy, Bovine Spongiform/classification , Encephalopathy, Bovine Spongiform/genetics , Prions/genetics , Prions/metabolism , Animals , Brain Chemistry , Cattle , Encephalopathy, Bovine Spongiform/metabolism , Encephalopathy, Bovine Spongiform/pathology , Genetic Predisposition to Disease , Mice , Mice, Transgenic , Phenotype , Prions/chemistry , Sheep
11.
Emerg Infect Dis ; 18(12): 2028-31, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23171544

ABSTRACT

We compared transmission characteristics for prions from L-type bovine spongiform encephalopathy and MM2-cortical sporadic Creutzfeldt-Jakob disease in the Syrian golden hamster and an ovine prion protein-transgenic mouse line and isolated distinct prion strains. Our findings suggest the absence of a causal relationship between these diseases, but further investigation is warranted.


Subject(s)
Creutzfeldt-Jakob Syndrome/transmission , Encephalopathy, Bovine Spongiform/transmission , Prions/pathogenicity , Animals , Brain/metabolism , Brain/pathology , Cattle , Creutzfeldt-Jakob Syndrome/mortality , Creutzfeldt-Jakob Syndrome/pathology , Cricetinae , Encephalopathy, Bovine Spongiform/mortality , Encephalopathy, Bovine Spongiform/pathology , Humans , Mice , Mice, Transgenic , Prions/metabolism
12.
Emerg Infect Dis ; 18(1): 142-5, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22261009

ABSTRACT

We report transmission of atypical L-type bovine spongiform encephalopathy to mouse lemurs after oral or intracerebral inoculation with infected bovine brain tissue. After neurologic symptoms appeared, transmissibility of the disease by both inoculation routes was confirmed by detection of disease-associated prion protein in samples of brain tissue.


Subject(s)
Cheirogaleidae , Encephalopathy, Bovine Spongiform/transmission , Animal Feed , Animals , Brain/pathology , Cattle , Encephalopathy, Bovine Spongiform/classification , Female , Male , Prions/metabolism
13.
Neurobiol Aging ; 33(9): 2225-8, 2012 Sep.
Article in English | MEDLINE | ID: mdl-21813214

ABSTRACT

Our aim in this study was to investigate experimentally the possible in vivo transmission of a synucleinopathy, using a transgenic mouse model (TgM83) expressing the human A53T mutated α-synuclein. Brain homogenates from old TgM83 mice showing motor clinical signs due to the synucleinopathy and containing insoluble and phosphorylated (pSer129) α-synuclein were intracerebrally inoculated in young TgM83 mice. This triggered an early onset of characteristic motor clinical signs, compared with uninoculated TgM83 mice or to mice inoculated with a brain homogenate from a young, healthy TgM83 mouse. This early disease was associated with insoluble α-synuclein phosphorylated on Ser129, as already identified in old and sick uninoculated TgM83 transgenic mice. Although the molecular mechanisms remain to be determined, acceleration of the pathology following inoculation of mice expressing human mutated α-synuclein with tissues from mice affected by the synucleinopathy, could be consistent with "prion-like" propagation of the disease.


Subject(s)
Intermediate Filament Proteins/genetics , Mutation/genetics , Prion Diseases/genetics , Prion Diseases/metabolism , Age Factors , Animals , Disease Models, Animal , Humans , Intermediate Filament Proteins/metabolism , Mice , Mice, Inbred C57BL , Mice, Transgenic , Movement Disorders/etiology , Movement Disorders/genetics , Phosphorylation/genetics , Prion Diseases/complications , Prion Diseases/mortality , Serine/genetics , Survival Analysis
14.
J Neuropathol Exp Neurol ; 70(5): 377-85, 2011 May.
Article in English | MEDLINE | ID: mdl-21487306

ABSTRACT

There is a growing interest in the potential roles of misfolded protein interactions in neurodegeneration. To investigate this issue, we inoculated 3 prion strains intracerebrally into transgenic (TgM83) mice that overexpress human A53T α-synuclein. In comparison to nontransgenic controls, there was a striking decrease in the incubation periods of scrapie, classic and H-type bovine spongiform encephalopathies(C-BSE and H-BSE), with conservation of the histopathologic and biochemical features characterizing these 3 prion strains. TgM83 mice died of scrapie or C-BSE prion diseases before accumulating the insoluble and phosphorylated forms of α-synuclein specific to late stages of synucleinopathy. In contrast, the median incubation time for TgM83 mice inoculated with H-BSE was comparable to that observed when these mice were uninfected, thereby allowing the development of molecular alterations of α-synuclein. The last 4 mice of this cohort exhibited early accumulations of H-BSE prion protein along with α-synuclein pathology. The results indicate that a prion disease was triggered concomitantly with an overt synucleinopathy in some transgenic mice overexpressing human A53T α-synuclein after intracerebral inoculation with an H-BSE prion strain.


Subject(s)
Brain/pathology , Encephalopathy, Bovine Spongiform/pathology , Prions/genetics , Scrapie/pathology , alpha-Synuclein/genetics , Animals , Blotting, Western , Brain/metabolism , Cattle , Encephalopathy, Bovine Spongiform/genetics , Encephalopathy, Bovine Spongiform/transmission , Immunohistochemistry , Kaplan-Meier Estimate , Mice , Mice, Transgenic , Prions/metabolism , Scrapie/genetics , Scrapie/transmission , alpha-Synuclein/metabolism
15.
J Virol ; 85(4): 1906-8, 2011 Feb.
Article in English | MEDLINE | ID: mdl-21123380

ABSTRACT

We investigated the susceptibilities of Syrian golden hamsters to transmissible spongiform encephalopathy agents from cattle. We report efficient transmission of the L-type atypical bovine spongiform encephalopathy (BSE) agent into hamsters. Importantly, hamsters were also susceptible to the transmissible mink encephalopathy agent from cattle, which has molecular features similar to those of the L-type BSE agent, as also shown in bovinized transgenic mice. In sharp contrast, hamsters could not be infected with classical or H-type BSE agents from cattle. However, previous adaptation of the classical BSE agent in wild-type mice led to efficient transmission. Thus, this study demonstrates the existence of distinct "strain barriers" upon the transmission of bovine prions in hamsters.


Subject(s)
Encephalopathy, Bovine Spongiform/transmission , Mesocricetus/metabolism , Prions/pathogenicity , Animals , Brain/metabolism , Brain/pathology , Cattle , Cricetinae , Encephalopathy, Bovine Spongiform/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , PrPSc Proteins/genetics , PrPSc Proteins/metabolism , Prions/genetics , Prions/metabolism , Species Specificity
16.
J Gen Virol ; 91(Pt 2): 570-4, 2010 Feb.
Article in English | MEDLINE | ID: mdl-19828761

ABSTRACT

The cerebral prion protein (PrP) isolated in the absence of proteinase K digestion, from ruminants prion sources transmitted to ovine transgenic mice, was studied by Western blot analysis. A C2 PrP fragment, showing strain-specific cleavages, similar to those observed after proteinase K or thermolysin digestion, accumulated in the brain. 'CH1641-like' scrapie was characterized by the unique accumulation of a more C-terminally cleaved PrP fragment (CTF14). A similar, protease-resistant, PrP product was observed after proteinase K or thermolysin digestion. Whereas classical BSE appeared highly resistant to thermolysin digestion, CH1641 and 'CH1641-like' natural isolates did not show any remarkable feature regarding resistance to thermolysin. Thus, the molecular strain-specific features in the brain of transmissible spongiform encephalopathy infected mice essentially reflect the PrP proteolytic processing occurring in vivo.


Subject(s)
Disease Models, Animal , Encephalopathy, Bovine Spongiform/transmission , Mice , PrPC Proteins/metabolism , Scrapie/transmission , Animals , Brain/metabolism , Cattle , Encephalopathy, Bovine Spongiform/metabolism , Endopeptidase K/metabolism , Mice, Transgenic , Protein Processing, Post-Translational , Scrapie/metabolism , Sheep , Species Specificity , Thermolysin/metabolism
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