sirolimus and versipelostatin

We recently isolated a macrocyclic compound, versipelostatin (VST), that exerts in vivo antitumor activity. VST shows unique, selective cytotoxicity to glucose-deprived tumor cells by preventing the unfolded protein response (UPR). Here we show that eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), a negative regulator of eukaryotic initiation factor 4E-mediated protein translation, plays a role in the UPR-inhibitory action of VST. Indeed, 4E-BP1 is aberrantly activated by VST. This activation occurs specifically during glucose deprivation and results in profound translation repression and prevents induction of the typical UPR markers glucose-regulated protein (GRP) 78 and activating transcription factor (ATF) 4. Our overexpression and knockdown experiments showed that 4E-BP1 can regulate GRP78 and ATF4 expression. These mechanisms appear to be specific for VST. By contrast, rapamycin, which activates 4E-BP1 regardless of cellular glucose availability, has only marginal effects on the expression of GRP78 and ATF4. Our present findings demonstrate that aberrant 4E-BP1 activation can contribute to UPR preventing by VST, possibly through a mechanism that does not operate in rapamycin-treated cells.

Topics: Activating Transcription Factor 4; Adaptor Proteins, Signal Transducing; Apoptosis; Blotting, Western; Cell Cycle Proteins; Cell Proliferation; Cells, Cultured; Endoplasmic Reticulum Chaperone BiP; Gene Expression Regulation; Heat-Shock Proteins; Humans; Immunosuppressive Agents; Luciferases; Macrolides; Oligosaccharides; Phosphoproteins; Phosphorylation; Protein Biosynthesis; RNA, Small Interfering; Sirolimus; Unfolded Protein Response