The interplay between a GC-rich oligonucleotide and copper ions on prion protein conformational and phase transitions

Int J Biol Macromol. 2021 Mar 15:173:34-43. doi: 10.1016/j.ijbiomac.2021.01.097. Epub 2021 Jan 18.

Abstract

The prion protein (PrP) misfolding to its infectious form is critical to the development of prion diseases, whereby various ligands are suggested to participate, such as copper and nucleic acids (NA). The PrP globular domain was shown to undergo NA-driven liquid-liquid phase separation (LLPS); this latter may precede pathological aggregation. Since Cu(II) is a physiological ligand of PrP, we argue whether it modulates phase separation altogether with nucleic acids. Using recombinant PrP, we investigate the effects of Cu(II) (at 6 M equivalents) and a previously described PrP-binding GC-rich DNA (equimolarly to protein) on PrP conformation, oligomerization, and phase transitions using a range of biophysical techniques. Raman spectroscopy data reveals the formation of the ternary complex. Microscopy suggests that phase separation is mainly driven by DNA, whereas Cu(II) has no influence. Our results show that DNA can be an adjuvant, leading to the structural conversion of PrP, even in the presence of an endogenous ligand, copper. These results provide new insights into the role of Cu(II) and NA on the phase separation, structural conversion, and aggregation of PrP, which are critical events leading to neurodegeneration.

Keywords: Aggregation; Conformational transition; Copper; Nucleic acid; Phase separation; Prion; Spectroscopy.

MeSH terms

  • Animals
  • Cations, Divalent
  • Cloning, Molecular
  • Copper / chemistry*
  • Copper / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism
  • Mice
  • Oligonucleotides / chemistry*
  • Oligonucleotides / genetics
  • Oligonucleotides / metabolism
  • Pregnancy Proteins / chemistry*
  • Pregnancy Proteins / genetics
  • Pregnancy Proteins / metabolism
  • Protein Aggregates*
  • Protein Conformation
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Cations, Divalent
  • Oligonucleotides
  • Plfr protein, mouse
  • Pregnancy Proteins
  • Protein Aggregates
  • Recombinant Proteins
  • Copper
  • cupric chloride