Exp Mol Med.  2016 Sep;48(9):e260. 10.1038/emm.2016.84.

ERK-mediated phosphorylation of BIS regulates nuclear translocation of HSF1 under oxidative stress

• Kim HY
• Kim YS
• Yun HH
• Im CN
• Ko JH
• Lee JH

• KMID: 2361547
• DOI: http://doi.org/10.1038/emm.2016.84

Abstract

B-cell lymphoma (BCL)-2-interacting cell death suppressor (BIS) has diverse cellular functions depending on its binding partners. However, little is known about the effects of biochemical modification of BIS on its various activities under oxidative stress conditions. In this study, we showed that Hâ‚‚Oâ‚‚ reduced BIS mobility on SDS-polyacrylamide gels in a time-dependent manner via the activation of extracellular signaling-regulated kinase (ERK). The combined results of mass spectroscopy and computational prediction identified Thr285 and Ser289 in BIS as candidate residues for phosphorylation by ERK under oxidative stress conditions. Deletion of these sites resulted in a partial reduction in the Hâ‚‚Oâ‚‚-induced mobility shift relative to that of the wild-type BIS protein; overexpression of the deletion mutant sensitized A172 cells to Hâ‚‚Oâ‚‚-induced cell death without increasing the level of intracellular reactive oxygen species. Expression of the BIS deletion mutant decreased the level of heat shock protein (HSP) 70 mRNA following Hâ‚‚Oâ‚‚ treatment, which was accompanied by impaired nuclear translocation of heat shock transcription factor (HSF) 1. Co-immunoprecipitation assays revealed that the binding of wild-type BIS to HSF1 was decreased by oxidative stress, while the binding of the BIS deletion mutant to HSF1 was not affected. These results indicate that ERK-dependent phosphorylation of BIS has a role in the regulation of nuclear translocation of HSF1 likely through modulation of its interaction affinity with HSF1, which affects HSP70 expression and sensitivity to oxidative stress.

MeSH Terms

Cell Death
Gels
Heat-Shock Proteins
Hot Temperature
Immunoprecipitation
Lymphoma, B-Cell
Mass Spectrometry
Oxidative Stress*
Phosphorylation*
Phosphotransferases
Reactive Oxygen Species
RNA, Messenger
Shock
Transcription Factors
Gels
Heat-Shock Proteins
Phosphotransferases
RNA, Messenger
Reactive Oxygen Species
Transcription Factors