Manganese-induced downregulation of astroglial glutamine transporter SNAT3 involves ubiquitin-mediated proteolytic system

Marta Sidoryk-Wecgrzynowicz, Eun Sook Lee, Mingwei Ni, Michael Aschner

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

SNAT3 is a major facilitator of glutamine (Gln) efflux from astrocytes, supplying Gln to neurons for neurotransmitter synthesis. Our previous investigations have shown that, in primary cortical astrocyte cultures, SNAT3 protein is degraded after exposure to manganese (Mn2+). The present studies were performed to identify the processes responsible for this effect. One of the well-established mechanisms for protein-level regulation is posttranslational modification via ubiquitination, which leads to the rapid degradation of proteins by the 26S proteasome pathway. Here, we show that astrocytic SNAT3 directly interacts with the ubiquitin ligase, Nedd4-2 (neural precursor cells expressed developmentally downregulated 4-2), and that Mn2+ increases both Nedd4-2 mRNA and protein levels. Additionally, we have found that Mn2+ exposure elevates astrocytic ubiquitin B mRNA expression, free ubiquitin protein levels, and total protein ubiquitination. Furthermore, Mn2+ effectively decreases astrocytic mRNA expression and the phosphorylation of serum and glucocorticoid-inducible kinase, a regulatory protein, which, in the active phosphorylated form, is responsible for the phosphorylation and subsequent inactivation of Nedd4-2. Additional findings establish that Mn2+ increases astrocytic caspase-like proteolytic proteasome activity and that the Mn2+-dependent degradation of SNAT3 protein is blocked by the proteasome inhibitors, N-acetylleu-leu-norleucinal and lactacystin. Combined, these results demonstrate that Mn2+-induced SNAT3 protein degradation and the dysregulation of Gln homeostasis in primary astrocyte cultures proceeds through the ubiquitin-mediated proteolytic system.

Original languageEnglish (US)
Pages (from-to)1905-1912
Number of pages8
JournalGlia
Volume58
Issue number16
DOIs
StatePublished - Dec 2010
Externally publishedYes

Keywords

  • Glutamine
  • Manganese
  • Nedd4-2/SGK1 signaling
  • Proteasome
  • SNAT3
  • Ubiquitination

ASJC Scopus subject areas

  • Neurology
  • Cellular and Molecular Neuroscience

Fingerprint Dive into the research topics of 'Manganese-induced downregulation of astroglial glutamine transporter SNAT3 involves ubiquitin-mediated proteolytic system'. Together they form a unique fingerprint.

  • Cite this