Further analysis of nucleic acids in purified scrapie prion preparations by improved return refocusing gel electrophoresis

Kellings, Klaus; Meyer, Norbert; Mirenda, C.; Prusiner, Stanley B.; Riesner, Detlev

doi:10.1099/0022-1317-73-4-1025

Cited by 118 publications

(69 citation statements)

References 11 publications

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“…These experiments show that the infectious agent is physically associated with PrP sc or a molecule very similar to it, but do not preclude the association of the PrP-derived molecule with another component. It has, however, been shown that highly purified prion preparations contain less than one molecule of nucleic acid larger than about 100 nucleotides [33].…”

Section: Physical Linkage Of Prp Sc and Prionsmentioning

confidence: 99%

Molecular biology of transmissible spongiform encephalopathies

1996

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The prion, the transmissible agent that causes spongiform encephalopathies such as serapie, bovine spongiform encephalopathy (BSE) and Creutzfeldt-Jakob disease, is believed to be devoid of nucleic acid and identical with PrP so, a modified form of the normal host protein PrP ¢ which is encoded by the single cop~v gene Prnp. The 'protein only" hypothesis proposes that PrP ~c, when introduced into a normal host, causes the conversion of PrP c into prpS~; it therefore predicts that an animal devoid of PrP c should be resistant to prion diseases. We generated homozygous Prnp °1° ('PrP knockout') mice and showed that, after inoculation with prions, they remained free of scrapie for at least 2 years while wild-type controls all died within 6 months. There was no propagation of prions in the Prnp °t° animals. Surprisingly, heterozygous Prnp °t+ mice, which express PrP c at about half the normal level, also showed enhanced resistance to scrapie disease despite high levels of infectious agent and PrP ~c in the brain early on. After introduction of murine PrP transgenes Prnp °t° mice became highly susceptible to mouse but not to hamster prions, while the insertion of Syrian hamster PrP transgenes rendered them susceptible to hamster but to a much lesser extent to mouse prions. These complementation experiments paved the way to the application of reverse genetics. We have prepared animals transgenic for genes encoding PrP with amino terminal deletions of various lengths and have found that PrP lacking 48 amino proximal amino acids, which comprise four of the five octa repeats of PrP, is still biologically active.

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Section: Physical Linkage Of Prp Sc and Prionsmentioning

confidence: 99%

Molecular biology of transmissible spongiform encephalopathies

1996

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“…These results, and numerous unsuccessful attempts to detect a scrapiespecific nucleic acid (36)(37)(38)(39)(40), militate against a foreign nucleic acid as an essential component of the "infectious" prion, despite proposals to the contrary (32, [41][42][43] …”

Section: Methodsmentioning

confidence: 99%

Serial transmission in rodents of neurodegeneration from transgenic mice expressing mutant prion protein.

Hsiao

Groth

Scott

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

259

134

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“…The key molecular event in prion diseases is the conformational change of a host-encoded prion protein, denoted as PrP C , into the disease-causing isoform, PrP Sc (1). Because prions do not contain any genetic information in the form of nucleic acid (2,3), the information for prions is enciphered in the structure of the pathological isoform. Prion replication occurs by converting PrP C to PrP Sc ; the pool of PrP C is replenished by the cellular synthesis of PrP C .…”

Section: P Rion Diseases Are Fatal Neurodegenerative Diseases That Imentioning

confidence: 99%

Mechanisms of prion protein assembly into amyloid

Stöhr

Weinmann

Wille

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

132

112

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The conversion of the ␣-helical, cellular isoform of the prion protein (PrP C ) to the insoluble, ␤-sheet-rich, infectious, diseasecausing isoform (PrP Sc ) is the key event in prion diseases. In an earlier study, several forms of PrP were converted into a fibrillar state by using an in vitro conversion system consisting of low concentrations of SDS and 250 mM NaCl. Here, we characterize the structure of the fibril precursor state, that is, the soluble state under fibrillization conditions. CD spectroscopy, analytical ultracentrifugation, and chemical cross-linking indicate that the precursor state exists in a monomer-dimer equilibrium of partially denatured, ␣-helical PrP, with a well defined contact site of the subunits in the dimer. Using fluorescence with thioflavin T, we monitored and quantitatively described the kinetics of seeded fibril formation, including dependence of the reaction on substrate and seed concentrations. Exponential, seed-enhanced growth can be achieved in homogeneous solution, which can be enhanced by sonication. From these data, we propose a mechanistic model of fibrillization, including the presence of several intermediate structures. These studies also provide a simplified amplification system for prions.dimer ͉ seeding ͉ fibril ͉ precursor state

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Further analysis of nucleic acids in purified scrapie prion preparations by improved return refocusing gel electrophoresis

Cited by 118 publications

References 11 publications

Molecular biology of transmissible spongiform encephalopathies

Molecular biology of transmissible spongiform encephalopathies

Serial transmission in rodents of neurodegeneration from transgenic mice expressing mutant prion protein.

Mechanisms of prion protein assembly into amyloid

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