Mechanism of zinc-induced phosphorylation of p70 S6 kinase and glycogen synthase kinase 3beta in SH-SY5Y neuroblastoma cells

J Neurochem. 2005 Mar;92(5):1104-15. doi: 10.1111/j.1471-4159.2004.02948.x.

Abstract

We have previously reported an aberrant accumulation of activated protein kinase B (PKB), glycogen synthase kinase (GSK)-3beta, extracellular signal-regulated kinase (ERK1/2), c-Jun N-terminal kinase (JNK), p38 and p70 S6 kinase (p70S6K) in neurons bearing neurofibrillary tangles (NFTs) in Alzheimer's disease (AD). However, the mechanism by which these tau candidate kinases are involved in the regulation of p70S6K and GSK-3beta phosphorylation is unknown. In the current study, 100 microM zinc sulfate was used, and influences of various components of phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) pathways on p70S6K and GSK-3beta phosphorylation have been investigated in serum-deprived SH-SY5Y neuroblastoma cells. We found that zinc could induce an increase of phosphorylated (p) p70S6K, p-PKB, p-GSK-3beta, p-ERK1/2, p-JNK and p-p38, especially in long-term treatment (4-8 h). Treatment with different inhibitors including rapamycin, wortmannin, LY294002, and U0126, and their combinations, indicated that phosphorylation of p70S6K and GSK-3beta is regulated by rapamycin-dependent, PI3K and MAPK pathways. Furthermore, phosphorylation of p70S6K and GSK-3beta affected levels of tau unphosphorylated at the Tau-1 site and phosphorylated at the PHF-1 site, and p70S6K phosphorylation affected the total tau level. Thus, 100 microM zinc might activate PKB, GSK-3beta, ERK1/2, JNK, p38 and p70S6K, that are consequently involved in tau changes in SH-SY5Y cells.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Animals
  • Antibiotics, Antineoplastic / pharmacology
  • Blotting, Western / methods
  • Brain / drug effects
  • Brain / metabolism
  • Brain / ultrastructure
  • Cell Line, Tumor / drug effects*
  • Cell Survival / drug effects
  • Culture Media, Serum-Free / pharmacology
  • Cytoplasm / drug effects
  • Cytoplasm / metabolism
  • Cytoplasm / ultrastructure
  • Dose-Response Relationship, Drug
  • Drug Interactions
  • Enzyme Inhibitors / pharmacology
  • Glycogen Synthase Kinase 3 / antagonists & inhibitors
  • Glycogen Synthase Kinase 3 / metabolism*
  • Glycogen Synthase Kinase 3 beta
  • Humans
  • Immunohistochemistry / methods
  • In Vitro Techniques
  • Microscopy, Immunoelectron / methods
  • Models, Biological
  • Neuroblastoma
  • Phosphorylation / drug effects
  • Rats
  • Ribosomal Protein S6 Kinases, 70-kDa / antagonists & inhibitors
  • Ribosomal Protein S6 Kinases, 70-kDa / metabolism*
  • Sirolimus / pharmacology
  • Tetrazolium Salts
  • Thiazoles
  • Time Factors
  • Zinc / pharmacology*
  • Zinc Sulfate / pharmacology
  • rab5 GTP-Binding Proteins / metabolism
  • tau Proteins / metabolism

Substances

  • Antibiotics, Antineoplastic
  • Culture Media, Serum-Free
  • Enzyme Inhibitors
  • Tetrazolium Salts
  • Thiazoles
  • tau Proteins
  • Zinc Sulfate
  • GSK3B protein, human
  • Glycogen Synthase Kinase 3 beta
  • Gsk3b protein, rat
  • Ribosomal Protein S6 Kinases, 70-kDa
  • Glycogen Synthase Kinase 3
  • rab5 GTP-Binding Proteins
  • thiazolyl blue
  • Zinc
  • Sirolimus