Overexpression of PLK3 Mediates the Degradation of Abnormal Prion Proteins Dependent on Chaperone-Mediated Autophagy

Mol Neurobiol. 2017 Aug;54(6):4401-4413. doi: 10.1007/s12035-016-9985-0. Epub 2016 Jun 25.

Abstract

Polo-like kinase 3 (PLK3) is the main cause of cell cycle reentry-related neuronal apoptosis which has been implicated in the pathogenesis of prion diseases. Previous work also showed the regulatory activity of exogenous PLK3 on the degradation of PrP (prion protein) mutants and pathogenic PrPSc; however, the precise mechanisms remain unknown. In this study, we identified that the overexpression of PLK3-mediated degradation of PrP mutant and PrPSc was repressed by lysosome rather than by proteasomal and macroautophagy inhibitors. Core components of chaperone-mediated autophagy (CMA) effectors, lysosome-associated membrane protein type 2A (LAMP2a), and heat shock cognate protein 70 (Hsc70) are markedly decreased in the HEK293T cells expressing PrP mutant and scrapie-infected cell line SMB-S15. Meanwhile, PrP mutant showed ability to interact with LAMP2a and Hsc70. Overexpression of PLK3 sufficiently increased the cellular levels of LAMP2a and Hsc70, accompanying with declining the accumulations of PrP mutant and PrPSc. The kinase domain (KD) of PLK3 was responsible for elevating LAMP2a and Hsc70. Knockdown of endogenous PLK3 enhanced the activity of macroautophagy in the cultured cells. Moreover, time-dependent reductions of LAMP2a and Hsc70 were also observed in the brain tissues of hamster-adapted scrapie agent 263K-infected hamsters, indicating an impairment of CMA during prion infection. Those data indicate that the overexpression of PLK3-mediated degradation of abnormal PrP is largely dependent on CMA pathway.

Keywords: Chaperone-mediated autophagy; Hsc70; LAMP2a; Lysosome; PLK3; Prion disease.

MeSH terms

  • Animals
  • Autophagy*
  • Brain / metabolism
  • Brain / pathology
  • Cricetinae
  • Down-Regulation
  • Gene Knockdown Techniques
  • HEK293 Cells
  • HSC70 Heat-Shock Proteins / metabolism
  • Humans
  • Lysosomal-Associated Membrane Protein 2 / metabolism
  • Lysosomes / metabolism
  • Molecular Chaperones / metabolism*
  • Mutant Proteins / metabolism
  • Prion Proteins / metabolism*
  • Protein Serine-Threonine Kinases / metabolism*
  • Proteolysis*
  • Scrapie / metabolism
  • Tumor Suppressor Proteins

Substances

  • HSC70 Heat-Shock Proteins
  • Lysosomal-Associated Membrane Protein 2
  • Molecular Chaperones
  • Mutant Proteins
  • Prion Proteins
  • Tumor Suppressor Proteins
  • PLK3 protein, human
  • Protein Serine-Threonine Kinases