Post-translational changes to PrP alter transmissible spongiform encephalopathy strain properties

Cancellotti, Enrico; Mahal, Sukhvir P.; Somerville, Robert A.; Diack, Abigail B.; Brown, Deborah; Piccardo, Pedro; Weissmann, Charles; Manson, Jean

doi:10.1038/emboj.2013.6

Cited by 57 publications

(54 citation statements)

References 67 publications

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“…Mature PrP C is rich in the diglycosylated form, whereas the glycoform ratio of PrP Sc is known to vary among strains (42)(43)(44). Studies using PrP glycan-lacking Tg mice revealed that the strain-specific characteristics of strain 79A were affected by the glycosylation status of PrP C , but those of strains ME7 and 301C were not (45). Meanwhile, enzymatic deglycosylation of PrP C failed to affect strain-specific pathological changes in serial PMCA experiments seeded with two murine strains, RML and 301C (46).…”

Section: Discussionmentioning

confidence: 99%

Conformational Properties of Prion Strains Can Be Transmitted to Recombinant Prion Protein Fibrils in Real-Time Quaking-Induced Conversion

Sano

Atarashi

Ishibashi

et al. 2014

J Virol

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The phenomenon of prion strains with distinct biological characteristics has been hypothesized to be involved in the structural diversity of abnormal prion protein (PrP Sc ). However, the molecular basis of the transmission of strain properties remains poorly understood. Real-time quaking-induced conversion (RT-QUIC) is a cell-free system that uses Escherichia coli-derived recombinant PrP (rPrP) for the sensitive detection of PrP Sc . To investigate whether the properties of various prion strains can be transmitted to amyloid fibrils consisting of rPrP (rPrP fibrils) using RT-QUIC, we examined the secondary structure, conformational stability, and infectivity of rPrP fibrils seeded with PrP Sc derived from either the Chandler or the 22L strain. In the first round of the reaction, there were differences in the secondary structures, especially in bands attributed to ␤-sheets, as determined by infrared spectroscopy, and conformational stability between Chandler-seeded (1st-rPrP-fib Ch ) and 22L-seeded (1st-rPrP-fib 22L ) rPrP fibrils. Of note, specific identifying characteristics of the two rPrP fibril types seen in the ␤-sheets resembled those of the original PrP Sc . Furthermore, the conformational stability of 1st-rPrP-fib Ch was significantly higher than that of 1st-rPrP-fib 22L , as with Chandler and 22L PrP Sc . The survival periods of mice inoculated with 1st-rPrP-fib Ch or 1st-rPrP-fib 22L were significantly shorter than those of mice inoculated with mixtures from the mock 1st-round RT-QUIC procedure. In contrast, these biochemical characteristics were no longer evident in subsequent rounds, suggesting that nonspecific uninfected rPrP fibrils became predominant probably because of their high growth rate. Together, these findings show that at least some strainspecific conformational properties can be transmitted to rPrP fibrils and unknown cofactors or environmental conditions may be required for further conservation. IMPORTANCEThe phenomenon of prion strains with distinct biological characteristics is assumed to result from the conformational variations in the abnormal prion protein (PrP Sc ). However, important questions remain about the mechanistic relationship between the conformational differences and the strain diversity, including how strain-specific conformations are transmitted. In this study, we investigated whether the properties of diverse prion strains can be transmitted to amyloid fibrils consisting of E. coli-derived recombinant PrP (rPrP) generated by real-time quaking-induced conversion (RT-QUIC), a recently developed in vitro PrP Sc formation method. We demonstrate that at least some of the strain-specific conformational properties can be transmitted to rPrP fibrils in the first round of RT-QUIC by examining the secondary structure, conformational stability, and infectivity of rPrP fibrils seeded with PrP Sc derived from either the Chandler or the 22L prion strain. We believe that these findings will advance our understanding of the conformational basis underlying prion strain diver...

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Section: Discussionmentioning

confidence: 99%

Conformational Properties of Prion Strains Can Be Transmitted to Recombinant Prion Protein Fibrils in Real-Time Quaking-Induced Conversion

Sano

Atarashi

Ishibashi

et al. 2014

J Virol

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“…In other models of prion infections, the glycosylation pattern of the host PrP has been shown to affect pathogenesis of specific prion strains (31,32,(61)(62)(63), and in cross-species experiments in vitro, PrPsen with less glycosylation bound to heterologous PrPres more readily than did PrPsen with larger amounts of glycosylation (34,44). Anchorless PrPres in the GPIneg-22L inoculum is mostly unglycosylated with some monoglycosylated PrPres, while the C57-22L inoculum contained mostly mono-and diglycosylated PrPres (36).…”

Section: Figmentioning

confidence: 99%

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

Race

Phillips

Meade-White

et al. 2015

J Virol

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Prion 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. Previous in vivo studies 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. IMPORTANCE Prion 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. P rion diseases, also known as transmissible spongiform encephalopathies (TSEs), are fatal neurodegenerative diseases that occur in many mammals, including humans. Central to all prion diseases is the conversion of the normal host prion protein (PrPsen or PrPC) into an abnormal, protease-resistant form (PrPres) associated with disease. In previous studies, PrPres and infectivity were generated concurrently in a cell-free in vitro system, proving that live cells were not required for formation of new prion infectivity in a cell-free in vitro system (1-4), a finding which supported the role of PrPres as the infectious prion agent (5). During the course of prion disease, repeated conversion of P...

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“…Glycosylation plays an important role in promoting correct folding of PrP C , and differences in glycosylation patterns contribute toward prion strain specificity (41)(42)(43). Moreover, the glycosylation status of PrP C has also been found to influence transmission of prions from the periphery to the CNS (44).…”

Section: Monensin Does Not Alter the Conversion Of Prp In An In Vitromentioning

confidence: 99%

Stimulating the Release of Exosomes Increases the Intercellular Transfer of Prions

Guo

Bellingham

Hill

2016

Journal of Biological Chemistry

126

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Post-translational changes to PrP alter transmissible spongiform encephalopathy strain properties

Cited by 57 publications

References 67 publications

Conformational Properties of Prion Strains Can Be Transmitted to Recombinant Prion Protein Fibrils in Real-Time Quaking-Induced Conversion

Conformational Properties of Prion Strains Can Be Transmitted to Recombinant Prion Protein Fibrils in Real-Time Quaking-Induced Conversion

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

Stimulating the Release of Exosomes Increases the Intercellular Transfer of Prions

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