Mutant p53 aggregates into prion-like amyloid oligomers and fibrils: implications for cancer

J Biol Chem. 2012 Aug 10;287(33):28152-62. doi: 10.1074/jbc.M112.340638. Epub 2012 Jun 19.

Abstract

Over 50% of all human cancers lose p53 function. To evaluate the role of aggregation in cancer, we asked whether wild-type (WT) p53 and the hot-spot mutant R248Q could aggregate as amyloids under physiological conditions and whether the mutant could seed aggregation of the wild-type form. The central domains (p53C) of both constructs aggregated into a mixture of oligomers and fibrils. R248Q had a greater tendency to aggregate than WT p53. Full-length p53 aggregated into amyloid-like species that bound thioflavin T. The amyloid nature of the aggregates was demonstrated using x-ray diffraction, electron microscopy, FTIR, dynamic light scattering, cell viabilility assay, and anti-amyloid immunoassay. The x-ray diffraction pattern of the fibrillar aggregates was consistent with the typical conformation of cross β-sheet amyloid fibers with reflexions of 4.7 Å and 10 Å. A seed of R248Q p53C amyloid oligomers and fibrils accelerated the aggregation of WT p53C, a behavior typical of a prion. The R248Q mutant co-localized with amyloid-like species in a breast cancer sample, which further supported its prion-like effect. A tumor cell line containing mutant p53 also revealed massive aggregation of p53 in the nucleus. We conclude that aggregation of p53 into a mixture of oligomers and fibrils sequestrates the native protein into an inactive conformation that is typical of a prionoid. This prion-like behavior of oncogenic p53 mutants provides an explanation for the negative dominance effect and may serve as a potential target for cancer therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Amyloid / chemistry*
  • Amyloid / genetics
  • Amyloid / metabolism
  • Benzothiazoles
  • Humans
  • Mutation, Missense*
  • Neoplasms / chemistry*
  • Neoplasms / genetics
  • Neoplasms / metabolism
  • Neoplasms / therapy
  • Prions*
  • Protein Multimerization*
  • Protein Structure, Quaternary
  • Protein Structure, Secondary
  • Thiazoles / chemistry
  • Tumor Suppressor Protein p53 / chemistry*
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism
  • X-Ray Diffraction

Substances

  • Amyloid
  • Benzothiazoles
  • Prions
  • TP53 protein, human
  • Thiazoles
  • Tumor Suppressor Protein p53
  • thioflavin T