Rapid ex vivo reverse genetics identifies the essential determinants of prion protein toxicity

Brain Pathol. 2023 Mar;33(2):e13130. doi: 10.1111/bpa.13130. Epub 2022 Nov 3.

Abstract

The cellular prion protein PrPC mediates the neurotoxicity of prions and other protein aggregates through poorly understood mechanisms. Antibody-derived ligands against the globular domain of PrPC (GDL) can also initiate neurotoxicity by inducing an intramolecular R208 -H140 hydrogen bond ("H-latch") between the α2-α3 and β2-α2 loops of PrPC . Importantly, GDL that suppresses the H-latch prolong the life of prion-infected mice, suggesting that GDL toxicity and prion infections exploit convergent pathways. To define the structural underpinnings of these phenomena, we transduced 19 individual PrPC variants to PrPC -deficient cerebellar organotypic cultured slices using adenovirus-associated viral vectors (AAV). We report that GDL toxicity requires a single N-proximal cationic residue (K27 or R27 ) within PrPC . Alanine substitution of K27 also prevented the toxicity of PrPC mutants that induce Shmerling syndrome, a neurodegenerative disease that is suppressed by co-expression of wild-type PrPC . K27 may represent an actionable target for compounds aimed at preventing prion-related neurodegeneration.

Keywords: neurodegeneration; prion disease; reverse genetic.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies
  • Mice
  • Neurodegenerative Diseases*
  • Prion Diseases* / genetics
  • Prion Proteins / genetics
  • Prions* / genetics
  • Reverse Genetics

Substances

  • Prion Proteins
  • Prions
  • Antibodies