Adaptive responses to physical and inflammatory stressors are mediated by
transcription factors and
molecular chaperones. The
transcription factor heat shock factor 1 (HSF1) has been implicated in extending lifespan in part by increasing expression of heat shock response genes.
Pyrrolidine dithiocarbamate (
PDTC) is a small
thiol compound that exerts in vivo and in vitro anti-inflammatory properties through mechanisms that remain unclear. Here we report that
PDTC induced the release of monomeric HSF1 from the
molecular chaperone heat shock protein 90 (Hsp90), with concomitant increase in HSF1 trimer formation, translocation to the nucleus, and binding to promoter of target genes in human HepG2 cells.
siRNA-mediated silencing of HSF1 blocked BAG3 gene expression by
PDTC. The
protein levels of the co-chaperone BAG3 and its interaction partner Hsp72 were stimulated by
PDTC in a dose-dependent fashion, peaking at 6h. Inhibition of Hsp90 function by
geldanamycin derivatives and
novobiocin elicited a pattern of HSF1 activation and BAG3 expression that was similar to
PDTC.
Chromatin immunoprecipitation studies showed that
PDTC and the inhibitor 17-dimethylaminoethylamino-17-demethoxygeldanamycin enhanced the binding of HSF1 to the promoter of several target genes, including BAG3, HSPA1A, HSPA1B,
FKBP4, STIP1 and UBB. Cell treatment with
PDTC increased significantly the level of Hsp90α
thiol oxidation, a posttranslational modification known to inhibit its chaperone function. These results unravel a previously unrecognized mechanism by which
PDTC and related compounds could confer cellular protection against
inflammation through HSF1-induced expression of heat shock response genes.