Familial human prion diseases associated with prion protein mutations Y226X and G131V are transmissible to transgenic mice expressing human prion protein

AbstractTransmissible spongiform encephalopathies (TSEs) are fatal neurodegenerative disorders associated with the misfolding and aggregation of the human prion protein (huPrP). Despite efforts into investigating the process of huPrP aggregation, the mechanisms triggering its misfolding remain elusive. A number of TSE-associated mutations of huPrP have been identified, but their role at the onset and progression of prion diseases is unclear. Here we report the NMR assignments of the C-terminal globular domain of the wild type huPrP and the pathological mutant T183A. The differences in chemical shifts between the two variants reveal conformational alterations in some structural elements of the mutant, whereas the analyses of secondary shifts and random coil index provide indications on the putative mechanisms of misfolding of T183A huPrP.

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Unaltered susceptibility to BSE in transgenic mice expressing human prion protein

Nature ◽

10.1038/39116 ◽

1997 ◽

Vol 389 (6650) ◽

pp. 526-526 ◽

Cited By ~ 6

Author(s):

John Collinge ◽

Mark S. Palmer ◽

Katie C. L. Sidle ◽

Andrew F. Hill ◽

Ian Gowland ◽

...

Keyword(s):

Transgenic Mice ◽

Prion Protein ◽

Human Prion Protein

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Dimerization of the cellular prion protein inhibits propagation of scrapie prions

Journal of Biological Chemistry ◽

10.1074/jbc.ra117.000990 ◽

2018 ◽

Vol 293 (21) ◽

pp. 8020-8031 ◽

Cited By ~ 2

Author(s):

Anna D. Engelke ◽

Anika Gonsberg ◽

Simrika Thapa ◽

Sebastian Jung ◽

Sarah Ulbrich ◽

...

Keyword(s):

Transgenic Mice ◽

Prion Protein ◽

Conformational Transition ◽

Prion Diseases ◽

Neuroblastoma Cells ◽

Direct Interaction ◽

Cellular Prion Protein ◽

Dominant Negative ◽

Dimerization Domain ◽

Hydrophobic Domain

A central step in the pathogenesis of prion diseases is the conformational transition of the cellular prion protein (PrPC) into the scrapie isoform, denoted PrPSc. Studies in transgenic mice have indicated that this conversion requires a direct interaction between PrPC and PrPSc; however, insights into the underlying mechanisms are still missing. Interestingly, only a subfraction of PrPC is converted in scrapie-infected cells, suggesting that not all PrPC species are suitable substrates for the conversion. On the basis of the observation that PrPC can form homodimers under physiological conditions with the internal hydrophobic domain (HD) serving as a putative dimerization domain, we wondered whether PrP dimerization is involved in the formation of neurotoxic and/or infectious PrP conformers. Here, we analyzed the possible impact on dimerization of pathogenic mutations in the HD that induce a spontaneous neurodegenerative disease in transgenic mice. Similarly to wildtype (WT) PrPC, the neurotoxic variant PrP(AV3) formed homodimers as well as heterodimers with WTPrPC. Notably, forced PrP dimerization via an intermolecular disulfide bond did not interfere with its maturation and intracellular trafficking. Covalently linked PrP dimers were complex glycosylated, GPI-anchored, and sorted to the outer leaflet of the plasma membrane. However, forced PrPC dimerization completely blocked its conversion into PrPSc in chronically scrapie-infected mouse neuroblastoma cells. Moreover, PrPC dimers had a dominant-negative inhibition effect on the conversion of monomeric PrPC. Our findings suggest that PrPC monomers are the major substrates for PrPSc propagation and that it may be possible to halt prion formation by stabilizing PrPC dimers.

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Structural effects of the highly protective V127 polymorphism on human prion protein

Communications Biology ◽

10.1038/s42003-020-01126-6 ◽

2020 ◽

Vol 3 (1) ◽

Author(s):

Laszlo L. P. Hosszu ◽

Rebecca Conners ◽

Daljit Sangar ◽

Mark Batchelor ◽

Elizabeth B. Sawyer ◽

...

Keyword(s):

Prion Protein ◽

Prion Disease ◽

Structural Changes ◽

Prion Diseases ◽

Single Point ◽

Structural Basis ◽

Single Point Mutation ◽

Solution Dynamics ◽

Human Prion Protein ◽

Prion Transmission

AbstractPrion diseases, a group of incurable, lethal neurodegenerative disorders of mammals including humans, are caused by prions, assemblies of misfolded host prion protein (PrP). A single point mutation (G127V) in human PrP prevents prion disease, however the structural basis for its protective effect remains unknown. Here we show that the mutation alters and constrains the PrP backbone conformation preceding the PrP β-sheet, stabilising PrP dimer interactions by increasing intermolecular hydrogen bonding. It also markedly changes the solution dynamics of the β2-α2 loop, a region of PrP structure implicated in prion transmission and cross-species susceptibility. Both of these structural changes may affect access to protein conformers susceptible to prion formation and explain its profound effect on prion disease.

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Prion Protein Expression Differences in Microglia and Astroglia Influence Scrapie-Induced Neurodegeneration in the Retina and Brain of Transgenic Mice

Journal of Virology ◽

10.1128/jvi.00865-07 ◽

2007 ◽

Vol 81 (19) ◽

pp. 10340-10351 ◽

Cited By ~ 31

Author(s):

Lisa Kercher ◽

Cynthia Favara ◽

James F. Striebel ◽

Rachel LaCasse ◽

Bruce Chesebro

Keyword(s):

Transgenic Mice ◽

Prion Protein ◽

Microglial Activation ◽

Prion Diseases ◽

Cell Types ◽

Mouse Strains ◽

Activation Markers ◽

Activated Microglia ◽

Morphological Criteria ◽

Scrapie Infection

ABSTRACT Activated microglia and astroglia are known to be involved in a variety of neurodegenerative diseases, including prion diseases. In the present experiments, we studied activation of astroglia and microglia after intraocular scrapie infection in transgenic mice expressing prion protein (PrP) in multiple cell types (tg7 mice) or in neurons only (tgNSE mice). In this model, scrapie infection and protease-resistant PrP deposition occurs in the retinas of both strains of mice, but retinal degeneration is observed only in tg7 mice. Our results showed that the retinas of tg7 and tgNSE mice both had astroglial activation with increased chemokine expression during the course of infection. However, only tg7 retinas exhibited strong microglial activation compared to tgNSE retinas, which showed little microglial activation by biochemical or morphological criteria. Therefore, microglial PrP expression might be required for scrapie-induced retinal microglial activation and damage. Furthermore, microglial activation preceded retinal neurodegeneration in tg7 mice, suggesting that activated microglia might contribute to the degenerative process, rather than being a response to the damage. Surprisingly, brain differed from retina in that an altered profile of microglial activation markers was upregulated, and the profiles in the two mouse strains were indistinguishable. Microglial activation in the brain was associated with severe brain vacuolation and neurodegeneration, leading to death. Thus, retinal and brain microglia appeared to differ in their requirements for activation, suggesting that different activation pathways occur in the two tissues.

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Increased Infectivity of Anchorless Mouse Scrapie Prions in Transgenic Mice Overexpressing Human Prion Protein

Journal of Virology ◽

10.1128/jvi.00362-15 ◽

2015 ◽

Vol 89 (11) ◽

pp. 6022-6032 ◽

Cited By ~ 11

Author(s):

Brent Race ◽

Katie Phillips ◽

Kimberly Meade-White ◽

James Striebel ◽

Bruce Chesebro

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Transgenic Mice ◽

Prion Protein ◽

Prion Diseases ◽

Human Species ◽

Species Barrier ◽

Gpi Anchor ◽

Prion Infection ◽

Barrier Model

ABSTRACTPrion protein (PrP) is found in all mammals, mostly as a glycoprotein anchored to the plasma membrane by a C-terminal glycosylphosphatidylinositol (GPI) linkage. Following prion infection, host protease-sensitive prion protein (PrPsen or PrPC) is converted into an abnormal, disease-associated, protease-resistant form (PrPres). Biochemical characteristics, such as the PrP amino acid sequence, and posttranslational modifications, such as glycosylation and GPI anchoring, can affect the transmissibility of prions as well as the biochemical properties of the PrPres generated. Previousin vivostudies on the effects of GPI anchoring on prion infectivity have not examined cross-species transmission. In this study, we tested the effect of lack of GPI anchoring on a species barrier model using mice expressing human PrP. In this model, anchorless 22L prions derived from tg44 mice were more infectious than 22L prions derived from C57BL/10 mice when tested in tg66 transgenic mice, which expressed wild-type anchored human PrP at 8- to 16-fold above normal. Thus, the lack of the GPI anchor on the PrPres from tg44 mice appeared to reduce the effect of the mouse-human PrP species barrier. In contrast, neither source of prions induced disease in tgRM transgenic mice, which expressed human PrP at 2- to 4-fold above normal.IMPORTANCEPrion protein (PrP) is found in all mammals, usually attached to cells by an anchor molecule called GPI. Following prion infection, PrP is converted into a disease-associated form (PrPres). While most prion diseases are species specific, this finding is not consistent, and species barriers differ in strength. The amino acid sequence of PrP varies among species, and this variability affects prion species barriers. However, other PrP modifications, including glycosylation and GPI anchoring, may also influence cross-species infectivity. We studied the effect of PrP GPI anchoring using a mouse-to-human species barrier model. Experiments showed that prions produced by mice expressing only anchorless PrP were more infectious than prions produced in mice expressing anchored PrP. Thus, the lack of the GPI anchor on prions reduced the effect of the mouse-human species barrier. Our results suggest that prion diseases that produce higher levels of anchorless PrP may pose an increased risk for cross-species infection.

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Truncated Forms of the Human Prion Protein in Normal Brain and in Prion Diseases

Journal of Biological Chemistry ◽

10.1074/jbc.270.32.19173 ◽

1995 ◽

Vol 270 (32) ◽

pp. 19173-19180 ◽

Cited By ~ 350

Author(s):

Shu G. Chen ◽

David B. Teplow ◽

Piero Parchi ◽

Jan K. Teller ◽

Pierluigi Gambetti ◽

...

Keyword(s):

Prion Protein ◽

Prion Diseases ◽

Normal Brain ◽

Human Prion Protein

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Strain-specific proteolytic processing of the prion protein in prion diseases of ruminants transmitted in ovine transgenic mice

Journal of General Virology ◽

10.1099/vir.0.014464-0 ◽

2009 ◽

Vol 91 (2) ◽

pp. 570-574 ◽

Cited By ~ 8

Author(s):

S. Nicot ◽

T. G. M. Baron

Keyword(s):

Transgenic Mice ◽

Prion Protein ◽

Prion Diseases ◽

Proteolytic Processing

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Cryo-EM structure of disease-related prion fibrils provides insights into seeding barriers

10.1101/2021.08.10.455830 ◽

2021 ◽

Author(s):

Qiuye Li ◽

Christopher P. Jaroniec ◽

Witold K. Surewicz

Keyword(s):

High Resolution ◽

Prion Protein ◽

Prion Disease ◽

Amyloid Fibrils ◽

Prion Diseases ◽

Protein Aggregates ◽

Direct Support ◽

Human Prion Protein ◽

Structural Insight

One of the least understood aspects of prion diseases is the structure of infectious prion protein aggregates. Here we report a high-resolution cryo-EM structure of amyloid fibrils formed by human prion protein with Y145Stop mutation that is associated with a familial prion disease. This structural insight allows us not only to explain previous biochemical findings, but also provides direct support for the conformational adaptability model of prion transmissibility barriers.

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Prion peptide-mediated calcium level alteration governs neuronal cell damage through AMPK-autophagy flux

10.21203/rs.2.24353/v2 ◽

2020 ◽

Author(s):

Ji-Hong Moon ◽

Sang-Youel Park

Keyword(s):

Cell Death ◽

Intracellular Calcium ◽

Prion Protein ◽

Prion Diseases ◽

Critical Role ◽

Neuronal Cell ◽

Autophagic Cell Death ◽

Autophagy Flux ◽

Human Prion Protein ◽

Ampk Activity

Abstract Background The distinctive molecular structure of the prion protein, PrPsc, is established only in mammals with infectious prion diseases. Prion protein characterizes either the transmissible pathogen itself or a primary constituent of the disease. Our report suggested that prion-mediated neuronal cell death is triggered by the autophagy flux. However, the alteration of intracellular calcium levels, AMPK activity in prion models has not been described. This study is focused on the effect of the changes in intracellular calcium levels on AMPK/autophagy flux pathway and PrP (106–126)-induced neurotoxicity. Methods Western blot and Immunocytochemistry was used to detect AMPK and autophagy-related protein expression. Flow cytometry and a TdT-mediated biotin-16-dUTP nick-end labeling (TUNEL) assay were used to detect the percentage of apoptotic cells. Calcium measurement was employed using fluo-4 by confocal microscope. Results We examined the effect of calcium homeostasis alterations induced by human prion protein on the autophagy flux in neuronal cells. Treatment with human prion protein increased the intracellular calcium concentration and induced cell death in primary neurons as well as in a neuronal cell line. Using pharmacological inhibitors, we showed that the L-type calcium channel is involved in the cellular entry of calcium ions. Inhibition of calcium uptake prevented autophagic cell death and reduction in AMP-activated protein kinase (AMPK) activity induced by human prion protein. Conclusion Our data demonstrated that prion protein-mediated calcium inflow plays a pivotal role in prion protein-induced autophagic cell death, and reduction in AMPK activity in neurons. Altogether, our results suggest that calcium influx might play a critical role in neurodegenerative diseases, including prion diseases.

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