Antifungal activity of amiodarone is mediated by disruption of calcium homeostasis

Soma Sen Gupta, Van Khue Ton, Veronica Beaudry, Samuel Rulli, Kyle Cunningham, Rajini Rao

Research output: Contribution to journalArticle

Abstract

The antiarrhythmic drug amiodarone was recently demonstrated to have novel broad range fungicidal activity. We provide evidence that amiodarone toxicity is mediated by disruption of Ca2+ homeostasis in Saccharomyces cerevisiae. In mutants lacking calcineurin and various Ca2+ transporters, including pumps (Pmrl and Pmc1), channels (Cch1/Mid1 and Yvc1), and exchangers (Vcx1), amiodarone sensitivity correlates with cytoplasmic calcium overload. Measurements of cytosolic Ca2+ by aequorin luminescence demonstrate a biphasic response to amiodarone. An immediate and extensive calcium influx was observed that was dose-dependent and correlated with drug sensitivity. The second phase consisted of a sustained release of calcium from the vacuole via the calcium channel Yvc1 and was independent of extracellular Ca2+ entry. To uncover additional cellular pathways involved in amiodarone sensitivity, we conducted a genome-wide screen of nearly 5000 single-gene yeast deletion mutants. 36 yeast strains with amiodarone hypersensitivity were identified, including mutants in transporters (pmr1, pdr5, and vacuolar H+-ATPase), ergosterol biosynthesis (erg3, erg6, and erg24), intracellular trafficking (vps45 and rcy1), and signaling (ypk1 and ptc1). Of three mutants examined (vps45, vma3, and rcy1), all were found to have defective calcium homeostasis, supporting a correlation with amiodarone hypersensitivity. We show that low doses of amiodarone and an azole (miconazole, fluconazole) are strongly synergistic and exhibit potent fungicidal effects in combination. Our findings point to the potentially effective application of amiodarone as a novel antimycotic, particularly in combination with conventional antifungals.

Original languageEnglish (US)
Pages (from-to)28831-28839
Number of pages9
JournalJournal of Biological Chemistry
Volume278
Issue number31
DOIs
StatePublished - Aug 1 2003

Fingerprint

Amiodarone
Homeostasis
Calcium
Yeast
Hypersensitivity
Genes
Yeasts
Aequorin
Vacuolar Proton-Translocating ATPases
Miconazole
Ergosterol
Azoles
Fluconazole
Calcineurin
Anti-Arrhythmia Agents
Biosynthesis
Gene Deletion
Calcium Channels
Vacuoles
Luminescence

ASJC Scopus subject areas

  • Biochemistry

Cite this

Antifungal activity of amiodarone is mediated by disruption of calcium homeostasis. / Gupta, Soma Sen; Ton, Van Khue; Beaudry, Veronica; Rulli, Samuel; Cunningham, Kyle; Rao, Rajini.

In: Journal of Biological Chemistry, Vol. 278, No. 31, 01.08.2003, p. 28831-28839.

Research output: Contribution to journalArticle

Gupta, Soma Sen ; Ton, Van Khue ; Beaudry, Veronica ; Rulli, Samuel ; Cunningham, Kyle ; Rao, Rajini. / Antifungal activity of amiodarone is mediated by disruption of calcium homeostasis. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 31. pp. 28831-28839.
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