title: DNA converts cellular prion protein into the beta-sheet conformation and inhibits prion peptide aggregation.
abstract: The main hypothesis for prion diseases proposes that the cellular protein (PrP(C)) can be altered into a misfolded, beta-sheet-rich isoform (PrP(Sc)), which in most cases undergoes aggregation. In an organism infected with PrP(Sc), PrP(C) is converted into the beta-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 alpha-helical conformation (cellular isoform) into a soluble, beta-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.
authors.affiliation: Programa de Biologia Estrutural, Departamento de Bioquimica Médica, Instituto de Ciências Biomédicas and Centro Nacional de Ressonância Magnética Nuclear de Macromoléculas, Universidade Federal do Rio de Janeiro, Rio de Janeiro RJ 21941-590, Brazil.
authors.forename: Y
authors.initials: Y
authors.lastname: Cordeiro
authors.forename: F
authors.initials: F
authors.lastname: Machado
authors.forename: L
authors.initials: L
authors.lastname: Juliano
authors.forename: M A
authors.initials: MA
authors.lastname: Juliano
authors.forename: R R
authors.initials: RR
authors.lastname: Brentani
authors.forename: D
authors.initials: D
authors.lastname: Foguel
authors.forename: J L
authors.initials: JL
authors.lastname: Silva
mesh_terms: D000818:Animals; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000378:metabolism*; D018621:PrPC Proteins / Q000737:chemistry* / Q000378:metabolism; D018620:PrPSc Proteins / Q000737:chemistry* / Q000378:metabolism; D017096:Prion Diseases / Q000378:metabolism*; D011487:Protein Conformation; D020033:Protein Isoforms; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D014508:Urea / Q000737:chemistry
chemical_list: D004268:DNA-Binding Proteins; D009841:Oligonucleotides; D018621:PrPC Proteins; D018620:PrPSc Proteins; D020033:Protein Isoforms; D014508:Urea; D004247:DNA