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The Isothiocyanate, Sulforaphane, Alters the Acidification of the Vacuole to Trigger Yeast Cell Death
Author(s) -
Murphy Michael Dennis,
Tucker Douglass W.,
Wilcox Alexander,
Thomas Stacy K,
Laprade David,
Austriaco Nicanor
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.834.5
Subject(s) - vacuole , sulforaphane , yeast , saccharomyces cerevisiae , programmed cell death , microbiology and biotechnology , mutant , chemistry , biology , biochemistry , apoptosis , gene , cytoplasm
The isothiocyanate, sulforaphane (SFN), isolated from cruciferous vegetables is a potential chemotherapeutic agent. Recent studies in mammalian cells have suggested that SFN works by causing cell cycle arrest and/or initiating programmed cell death (PCD). In order to identify genes that may be involved in the eukaryotic cell's response to SFN, we initiated a genetic screen using the yeast knockout (YKO) library of the budding yeast, Saccharomyces cerevisiae, to identify loss of function mutants that are hypersensitive to SFN. We have identified 311 SFN‐sensitive and 11 SFN‐resistant mutants in yeast ORFs that had been previously linked to vacuolar acidification suggesting that SFN kills cells by altering the acidification of the yeast vacuole. Staining with BCECF‐AM confirmed that SFN makes yeast vacuoles more basic. Strikingly, cells cultured in lethal concentrations of benzyl isothiocyanate and phenethyl isothiocyanate did not exhibit changes in vacuole acidification, suggesting that this trigger of programmed cell death is unique to sulforaphane. [Our laboratory is supported by the following grants: NIGMS R15 GM094712, NSF MRI‐R2 0959354 and NIH Grant 2 P20 RR016457 to the Rhode Island INBRE Program]