Changing a single amino acid in the N-terminus of murine PrP alters TSE incubation time across three species barriers

Barron, Rona; Thomson, Val; Jamieson, Elizabeth R.; Melton, David W.; Ironside, James W.; Will, Robert; Manson, Jean

doi:10.1093/emboj/20.18.5070

Cited by 113 publications

(101 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…7). Such an approach could also be interesting for generating artificial PrP Sc molecules with amino acid changes that might be associated with altered scrapie strain characteristics in mice (8,46). Thus, this system may also help to better understand the role of PrP sequence alterations on the emergence of TSE strains with new disease phenotypes.…”

Section: Discussionmentioning

confidence: 99%

Scrapie Infection of Prion Protein-deficient Cell Line upon Ectopic Expression of Mutant Prion Proteins

Maas

Geissen

Groschup

et al. 2007

Journal of Biological Chemistry

View full text Add to dashboard Cite

Expression of the cellular prion protein (PrP C ) is crucial for susceptibility to prions. In vivo, ectopic expression of PrP C restores susceptibility to prions and transgenic mice that express heterologous PrP on a PrP knock-out background have been used extensively to study the role of PrP alterations for prion transmission and species barriers. Here we report that prion protein knock-out cells can be rendered permissive to scrapie infection by the ectopic expression of PrP. The system was used to study the influence of sheep PrP-specific residues in mouse PrP on the infection process with mouse adapted scrapie. These studies reveal several critical residues previously not associated with species barriers and demonstrate that amino acid residue alterations at positions known to have an impact on the susceptibility of sheep to sheep scrapie also drastically influence PrP Sc formation by mouse-adapted scrapie strain 22L. Furthermore, our data suggest that amino acid polymorphisms located on the outer surfaces of helix 2 and 3 drastically impact conversion efficiency. In conclusion, this system allows for the fast generation of mutant PrP Sc that is entirely composed of transgenic PrP and is, thus, ideally suited for testing if artificial PrP molecules can affect prion replication. Transmission of infectivity generated in HpL3-4 cells expressing altered PrP molecules to mice could also help to unravel the potential influence of mutant PrP Sc on host cell tropism and strain characteristics in vivo.Prion diseases or transmissible spongiform encephalopathies (TSEs) 3 are fatal neurological disorders such as Creutzfeldt-Jakob diseases in humans, scrapie in sheep and goats, and bovine spongiform encephalopathy (BSE). A key event in prion diseases is the conversion of the cellular, hostencoded prion protein (PrP C ) to its abnormal isoform PrP Since then transgenic animals have been used extensively to unravel the influence of specific PrP amino acid residues or domains on prion susceptibility (6 -11). Although these hallmark experiments added important information to prion biology (12), generation of transgenic animals is laborious and expensive and, thus, not suited for testing broad sets of PrP mutants.Alternative cell culture models for prion diseases have greatly helped us to understand the molecular mechanism of PrP Sc formation (13,14), the role of the PrP amino acid sequence for the TSE species barrier, and how PrP structural domains affect conversion of PrP C to PrP Sc (15)(16)(17)(18)(19)(20). However, so far comparative studies on the direct influence of PrP alterations on the prion infection process (as opposed to studies in persistently infected cell cultures) were mainly restricted to transgenic animal models (6,7,(21)(22)(23). One reason for this is that all so-far-identified cell lines susceptible to prions code for endogenous wild-type PrP. Therefore endogenous PrP might be co-expressed with the exogenous PrP of interest during infection experiments (24 -27). Thus, successful infection could still b...

show abstract

Section: Discussionmentioning

confidence: 99%

Scrapie Infection of Prion Protein-deficient Cell Line upon Ectopic Expression of Mutant Prion Proteins

Maas

Geissen

Groschup

et al. 2007

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…12 Transmission barriers are caused by amino acid sequence differences between the host cellular prion protein, PrP C , and the misfolded, aggregated conformation, PrP Sc . [13][14][15] The conformation of PrP Sc also plays a role in species barriers. For example, CWD prions are not transmissible to mice expressing human PrP C , yet are efficiently transmitted to mice expressing cervid PrP C , demonstrating the importance of amino acid sequence in susceptibility.…”

Section: Cross-species Cwd Prion Transmissionmentioning

confidence: 99%

Cross-species transmission of CWD prions

Kurt

Sigurdson

2016

Prion

View full text Add to dashboard Cite

“…9 In humans, disease susceptibility and phenotypic expression are influenced by PRNP mutations and by the polymorphism at codon 129 that encodes either methionine (M) or valine (V). [10][11][12] Distinct agent strains have been demonstrated [13][14][15][16] but have yet to be characterized in full.…”

mentioning

confidence: 99%

A refined method for molecular typing reveals that co-occurrence of PrPSc types in Creutzfeldt–Jakob disease is not the rule

Notari

Capellari

Langeveld

et al. 2007

Laboratory Investigation

View full text Add to dashboard Cite

Molecular typing in Creutzfeldt-Jakob disease (CJD) relies on the detection of distinct protease-resistant prion protein (PrP Sc ) core fragments, which differ in molecular mass or glycoform ratio. However, the definition and correct identification of CJD cases with a co-occurrence of PrP Sc types remains a challenge. With antibodies recognizing a linear epitope in the octapeptide repeat PrP region, supposed to distinguish between the two major PrP Sc isoforms (ie, types 1 and 2), it was recently shown that all type 2 cases display an associated band with a type 1 migration pattern, which led to the conclusion that multiple PrP Sc types regularly coexist in CJD. We studied brain samples from 53 sporadic CJD and 4 variant CJD subjects using a high-resolution electrophoresis, a wide range of proteinase K (PK) activities, the 'type 1-selective' antibody 12B2, and several unselective antibodies. We found that the type 1-like band detected by 12B2 in all CJD subtypes associated with PrP Sc type 2 is not a PK-resistant PrP Sc core but rather matches the physicochemical properties of partially cleaved fragments, which result from the several PK cleavage sites included in the N-terminal portion of PrP Sc . Furthermore, using gels with high resolution and a relatively high PK activity, we were able to increase the detection sensitivity of either type 1 or 2, when coexisting, to amount corresponding to 3-5% of the total PrP Sc signal (ie, weak band of one type/total PrP Sc ). Our results show that there are many pitfalls associated with the use of 'type 1 selective' antibodies for CJD typing studies and that co-occurrence of PrP Sc types in CJD is not the rule. The present results further validate the rationale basis of current CJD classification and the qualitative nature of molecular typing in CJD.

show abstract

Changing a single amino acid in the N-terminus of murine PrP alters TSE incubation time across three species barriers

Cited by 113 publications

References 60 publications

Scrapie Infection of Prion Protein-deficient Cell Line upon Ectopic Expression of Mutant Prion Proteins

Scrapie Infection of Prion Protein-deficient Cell Line upon Ectopic Expression of Mutant Prion Proteins

Cross-species transmission of CWD prions

A refined method for molecular typing reveals that co-occurrence of PrPSc types in Creutzfeldt–Jakob disease is not the rule

Contact Info

Product

Resources

About