Exp Mol Med.  2008 Dec;40(6):596-606. 10.3858/emm.2008.40.6.596.

Protein kinase C micron plays an essential role in hypertonicity-induced heat shock protein 70 expression

• Lim YS
• Lee JS
• Huang TQ
• Seo JS

• KMID: 1072940
• DOI: http://doi.org/10.3858/emm.2008.40.6.596

Abstract

Heat shock protein 70 (HSP70), which evidences important functions as a molecular chaperone and anti-apoptotic molecule, is substantially induced in cells exposed to a variety of stresses, including hypertonic stress, heavy metals, heat shock, and oxidative stress, and prevents cellular damage under these conditions. However, the molecular mechanism underlying the induction of HSP70 in response to hypertonicity has been characterized to a far lesser extent. In this study, we have investigated the cellular signaling pathway of HSP70 induction under hypertonic conditions. Initially, we applied a variety of kinase inhibitors to NIH3T3 cells that had been exposed to hypertonicity. The induction of HSP70 was suppressed specifically by treatment with protein kinase C (PKC) inhibitors (Go6976 and GF109203X). As hypertonicity dramatically increased the phosphorylation of PKC micron, we then evaluated the role of PKC micron in hypertonicity-induced HSP70 expression and cell viability. The depletion of PKC micron with siRNA or the inhibition of PKC micron activity with inhibitors resulted in a reduction in HSP70 induction and cell viability. Tonicity-responsive enhancer binding protein (TonEBP), a transcription factor for hypertonicity-induced HSP70 expression, was translocated rapidly into the nucleus and was modified gradually in the nucleus under hypertonic conditions. When we administered treatment with PKC inhibitors, the mobility shift of TonEBP was affected in the nucleus. However, PKC micron evidenced no subcellular co-localization with TonEBP during hypertonic exposure. From our results, we have concluded that PKC micron performs a critical function in hypertonicity-induced HSP70 induction, and finally cellular protection, via the indirect regulation of TonEBP modification.

Keyword

HSP70 heat-shock proteins; NFAT5 protein, human; protein kinase C; protein kinase inhibitors

MeSH Terms

Animals
Carbazoles/pharmacology
Cell Line
Flavonoids/pharmacology
HSP70 Heat-Shock Proteins/*biosynthesis
Humans
Indoles/pharmacology
Isoquinolines/pharmacology
MAP Kinase Signaling System/physiology
Maleimides/pharmacology
Mice
NFATC Transcription Factors/metabolism
Phosphorylation
Promoter Regions, Genetic
Protein Kinase C/antagonists & inhibitors/*physiology
Protein Transport
Saline Solution, Hypertonic/*pharmacology
Signal Transduction
Sulfonamides/pharmacology