The Prion Protein Has RNA Binding and Chaperoning Properties Characteristic of Nucleocapsid Protein NCp7 of HIV-1

Gabus, Caroline; Derrington, Edmund; Leblanc, Pascal; Chnaiderman, Jonás; Dormont, Dominique; Świętnicki, Wiesław; Morillas, Manuel; Surewicz, Witold K.; Marc, Daniel; Nandi, P. K.; Darlix, Jean Luc

doi:10.1074/jbc.m009754200

Cited by 175 publications

(188 citation statements)

References 50 publications

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“…DNA also binds to prion sequences containing the region that undergoes the transition to ␤-sheet, modulating their aggregation. Previous studies have shown that PrP may interact with sulfated glycans (27), DNA (28), and RNA (29). We also find that there is a dependence on the DNA sequence for binding to the murine prion protein.…”

supporting

confidence: 59%

DNA Converts Cellular Prion Protein into the β-Sheet Conformation and Inhibits Prion Peptide Aggregation

Cordeiro¹,

Machado²,

Juliano³

et al. 2001

Journal of Biological Chemistry

198

269

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The main hypothesis for prion diseases proposes that the cellular protein (PrP C ) can be altered into a misfolded, ␤-sheet-rich isoform (PrP Sc ), which in most cases undergoes aggregation. In an organism infected with PrP Sc , PrP C is converted into the ␤-sheet form, generating more PrP Sc . We find that sequence-specific DNA binding to recombinant murine prion protein (mPrP-(23-231)) converts it from an ␣-helical conformation (cellular isoform) into a soluble, ␤-sheet isoform similar to that found in the fibrillar state. The recombinant murine prion protein and prion domains bind with high affinity to DNA sequences. Several double-stranded DNA sequences in molar excess above 2:1 (pH 4.0) or 0.5:1 (pH 5.0) completely inhibit aggregation of prion peptides, as measured by light scattering, fluorescence, and circular dichroism spectroscopy. However, at a high concentration, fibers (or peptide aggregates) can rescue the peptide bound to the DNA, converting it to the aggregating form. Our results indicate that a macromolecular complex of prion-DNA may act as an intermediate for the formation of the growing fiber. We propose that host nucleic acid may modulate the delicate balance between the cellular and the misfolded conformations by reducing the protein mobility and by making the protein-protein interactions more likely. In our model, the infectious material would act as a seed to rescue the protein bound to nucleic acid. Accordingly, DNA would act on the one hand as a guardian of the Sc conformation, preventing its propagation, but on the other hand may catalyze Sc conversion and aggregation if a threshold level is exceeded.

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supporting

confidence: 59%

DNA Converts Cellular Prion Protein into the β-Sheet Conformation and Inhibits Prion Peptide Aggregation

Cordeiro¹,

Machado²,

Juliano³

et al. 2001

Journal of Biological Chemistry

198

269

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“…Further studies are required to distinguish between these possibilities and to identify the specific endogenous polyanions that facilitate prion propagation in vivo. Several classes of negatively charged macromolecules could potentially serve as cofactors for prion propagation, including: nucleic acids (17,(32)(33)(34)(35)(36)(37)(38), glycosaminoglycans (14-16), phospholipid-rich membranes (39)(40)(41)(42), and chaperone proteins (43). Interestingly, it has been proposed that polyanions could play a broad role in protein folding and misfolding, and the ability of polyanions to facilitate prion conversion may represent a specific example of that general concept (44).…”

Section: Discussionmentioning

confidence: 99%

Formation of native prions from minimal components in vitro

Deleault¹,

Harris²,

Rees³

et al. 2007

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

585

635

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The conformational change of a host protein, PrP C , into a disease-associated isoform, PrP Sc , appears to play a critical role in the pathogenesis of prion diseases such as Creutzfeldt–Jakob disease and scrapie. However, the fundamental mechanism by which infectious prions are produced in neurons remains unknown. To investigate the mechanism of prion formation biochemically, we conducted a series of experiments using the protein misfolding cyclic amplification (PMCA) technique with a preparation containing only native PrP C and copurified lipid molecules. These experiments showed that successful PMCA propagation of PrP Sc molecules in a purified system requires accessory polyanion molecules. In addition, we found that PrP Sc molecules could be formed de novo from these defined components in the absence of preexisting prions. Inoculation of samples containing either prion-seeded or spontaneously generated PrP Sc molecules into hamsters caused scrapie, which was transmissible on second passage. These results show that prions able to infect wild-type hamsters can be formed from a minimal set of components including native PrP C molecules, copurified lipid molecules, and a synthetic polyanion.

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“…These properties should be investigated further in order to gain more insight into the putative physiological role of PrP in RNA metabolism (25,26,28) and should provide ways to trap PrP in a disease-inactive conformation that might cleanse physiological fluids. …”

Section: Discussionmentioning

confidence: 99%

“…Large and small nucleic acids (21)(22)(23)(24) also interact with PrP. It has been shown that human, ovine, and murine PrPs possess nucleic acid binding and chaperoning activities similar to retroviral nucleocapsid protein (25,26), and these properties are associated with the unstructured N-terminal region of the protein (5,10). Previously reported 2Ј-OH aptamers raised against recombinant hamster prion protein bound to a site between residues 23-52 within this nonspecific nucleic acid binding region (27).…”

mentioning

confidence: 99%

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Characterization of 2′-Fluoro-RNA Aptamers That Bind Preferentially to Disease-associated Conformations of Prion Protein and Inhibit Conversion

Rhie

Kirby

Sayer

et al. 2003

Journal of Biological Chemistry

173

148

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We have isolated artificial ligands or aptamers for infectious prions in order to investigate conformational aspects of prion pathogenesis. The aptamers are 2 -fluoro-modified RNA produced by in vitro selection from a large, randomized library. One of these ligands (aptamer SAF-93) had more than 10-fold higher affinity for PrP Sc than for recombinant PrP C and inhibited the accumulation of PrP res in near physiological cell-free conversion assay. To understand the molecular basis of these properties and to distinguish specific from nonspecific aptamer-PrP interactions, we studied deletion mutants of bovine PrP in denatured, ␣-helix-rich and ␤-sheet-rich forms. We provide evidence that, like scrapie-associated fibrils (SAF), the ␤-oligomer of PrP bound to SAF-93 with at least 10-fold higher affinity than did the ␣-form. This differential affinity could be explained by the existence of two binding sites within the PrP molecule. Site 1 lies within residues 23-110 in the unstructured N terminus and is a nonspecific RNA binding site found in all forms of PrP. The region between residue 90 and 110 forms a hinge region that is occluded in the ␣-rich form of PrP but becomes exposed in the denatured form of PrP. Site 2 lies in the region C-terminal of residue 110. This site is ␤-sheet conformation-specific and is not recognized by control RNAs. Taken together, these data provide for the first time a specific ligand for a disease conformation-associated site in a region of PrP critical for conformational conversion. This aptamer could provide tools for the further analysis of the processes of PrP misfolding during prion disease and leads for the development of diagnostic and therapeutic approaches to TSEs.

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The Prion Protein Has RNA Binding and Chaperoning Properties Characteristic of Nucleocapsid Protein NCp7 of HIV-1

Cited by 175 publications

References 50 publications

DNA Converts Cellular Prion Protein into the β-Sheet Conformation and Inhibits Prion Peptide Aggregation

DNA Converts Cellular Prion Protein into the β-Sheet Conformation and Inhibits Prion Peptide Aggregation

Formation of native prions from minimal components in vitro

Characterization of 2′-Fluoro-RNA Aptamers That Bind Preferentially to Disease-associated Conformations of Prion Protein and Inhibit Conversion

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