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National Committee on Marine Sciences (NCMS)

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    Genomic and targeted approaches unveil the cell membrane as a major target of the antifungal cytotoxin amantelide A
    Elsadek, Lobna A.; Matthews, James H.; Nishimura, Shinichi; Nakatani, Takahiro; Ito, Airi; Gu, Tongjun; Luo, Danmeng; Salvador-Reyes, Lilibeth A.; Paul, Valerie J.; Kakeya, Hideaki; Luesch, Hendrik (Wiley, 2021-03-23)
    Amantelide A, a polyhydroxylated macrolide isolated from a marine cyanobacterium, displays broad-spectrum activity against mammalian cells, bacterial pathogens, and marine fungi. We conducted comprehensive mechanistic studies to identify the molecular targets and pathways affected by amantelide A. Our investigations relied on chemical structure similarities with compounds of known mechanisms, yeast knockout mutants, yeast chemogenomic profiling, and direct biochemical and biophysical methods. We established that amantelide A exerts its antifungal action by binding to ergosterol-containing membranes followed by pore formation and cell death, a mechanism partially shared with polyene antifungals. Binding assays demonstrated that amantelide A also binds to membranes containing epicholesterol or mammalian cholesterol, thus suggesting that the cytotoxicity to mammalian cells might be due to its affinity to cholesterol-containing membranes. However, membrane interactions were not completely dependent on sterols. Yeast chemogenomic profiling suggested additional direct or indirect effects on actin. Accordingly, we performed actin polymerization assays, which suggested that amantelide A also promotes actin polymerization in cell-free systems. However, the C-33 acetoxy derivative amantelide B showed a similar effect on actin dynamics in vitro but no significant activity against yeast. Overall, these studies suggest that the membrane effects are the most functionally relevant for amantelide A mechanism of action.
    This research was supported by the National Institutes of Health (grant R01CA172310 to H.L.), the Debbie and Sylvia DeSantis Chair Professorship (H.L), and a Grant-in Aid for Scientific Research (no. 17H06401 to S.N. and H.K.) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. L.E. was supported by the NIH/NIGMS T32GM136583 “Chemistry-Biology Interface Training Program at the University of Florida”. We thank Dr. Yanping Zhang (University of Florida) from the UF ICBR NextGen DNA Sequencing core facility for carrying out the nextgeneration sequencing, Dr. Maya Schuldiner (Weizmann Institute of Science, Israel) for providing the triple yeast deletion library, Dr. Kaoru Takegawa (Kyushu University, Japan) for providing the ergosterol mutants, Dr. Charles Boone laboratory (University of Toronto, Canada) for providing the wild-type S. cerevisiae Y7092 and Dr. Kalina Atanasova (CNPD3, University of Florida) for assistance with the imaging.
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    BFAR Administrative Circular No. 256: Series of 2015. Declaring malachite green and gentian violet as health hazards and prohibiting their use in food fish production and trade.
    (Bureau of Fisheries and Aquatic Resources, 2015-02-12)
    This Circular, consisting of 25 Sections divided into seven Chapters and one Annex, prohibits the use of malachite green and gentian violet in all stages of aquaculture production of fishery and aquatic products intended for human consumption and the distribution for commerce of fishery and aquatic products derived from aquaculture containing traces of the dyes. It shall be unlawful for any person to: 1) Use malachite green or gentian violet in the aquaculture production of food fish; 2) Produce, handle or process for sale, offer for sale, distribute in commerce fish or fishery product bearing or containing malachite green or gentian violet; and, 3) Import into the Philippines or export fish or fishery product bearing or containing malachite green or gentian violet. Any importer, exporter, aquaculture/farm operator, or distributor found violating any of the provisions of this Circular shall be subject to the following administrative sanctions: a) for the first offense, suspension for three months of the registration as importer, exporter, aquaculture/farm operator or distributor; b) for the second offense, six-month suspension of the registration; and, c) for the third offense, revocation of the registration and delisting.