title: Natamycin inhibits vacuole fusion at the priming phase via a specific interaction with ergosterol.
abstract: The antifungal antibiotic natamycin belongs to the family of polyene antibiotics. Its antifungal activity arises via a specific interaction with ergosterol in the plasma membrane (te Welscher et al., J. Biol. Chem. 283:6393-6401, 2008). However, this activity does not involve disruption of the membrane barrier function, a well-known property of other members of the polyene antibiotic family, such as filipin and nystatin. Here we tested the effect of natamycin on vacuole membrane fusion, which is known to be ergosterol dependent. Natamycin blocked the fusion of isolated vacuoles without compromising the barrier function of the vacuolar membrane. Sublethal doses of natamycin perturbed the cellular vacuole morphology, causing the formation of many more small vacuolar structures in yeast cells. Using vacuoles isolated from yeast strains deficient in the ergosterol biosynthesis pathway, we showed that the inhibitory activity of natamycin was dependent on the presence of specific chemical features in the structure of ergosterol that allow the binding of natamycin. We found that natamycin inhibited the priming stage of vacuole fusion. Similar results were obtained with nystatin. These results suggest a novel mode of action of natamycin and perhaps all polyene antibiotics, which involves the impairment of membrane fusion via perturbation of ergosterol-dependent priming reactions that precede membrane fusion, and they may point to an effect of natamycin on ergosterol-dependent protein function in general.
authors.affiliation: Department of Biochemistry of Membranes, Bijvoet Center, Institute of Biomembranes, Utrecht University, Utrecht CH, The Netherlands. y.m.tewelscher@uu.nl
authors.forename: Yvonne Maria
authors.initials: YM
authors.lastname: te Welscher
authors.forename: Lynden
authors.initials: L
authors.lastname: Jones
authors.forename: Martin Richard
authors.initials: MR
authors.lastname: van Leeuwen
authors.forename: Jan
authors.initials: J
authors.lastname: Dijksterhuis
authors.forename: Ben
authors.initials: B
authors.lastname: de Kruijff
authors.forename: Gary
authors.initials: G
authors.lastname: Eitzen
authors.forename: Eefjan
authors.initials: E
authors.lastname: Breukink
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology*; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004875:Ergosterol / Q000378:metabolism*; D005372:Filipin / Q000494:pharmacology; D017353:Gene Deletion; D005800:Genes, Fungal; D008561:Membrane Fusion / Q000187:drug effects; D008969:Molecular Sequence Data; D010866:Natamycin / Q000494:pharmacology*; D009761:Nystatin / Q000494:pharmacology; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010539:Permeability / Q000187:drug effects; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014617:Vacuoles / Q000187:drug effects / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D017931:DNA Primers; D004271:DNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; D009761:Nystatin; D005372:Filipin; D010866:Natamycin; D010088:Oxidoreductases; C039306:sterol delta-5 desaturase; C070590:ERG4 protein, S cerevisiae; D004875:Ergosterol
references.citation: Annu Rev Genet. 2000;34:255-295
references.pmid: 11092829
references.citation: J Biol Chem. 1985 Mar 10;260(5):2884-9
references.pmid: 3972808
references.citation: Science. 1999 Dec 17;286(5448):2361-4
references.pmid: 10600751
references.citation: Microbiol Mol Biol Rev. 2006 Mar;70(1):177-91
references.pmid: 16524922
references.citation: Biochim Biophys Acta. 1974 Feb 26;339(1):44-56
references.pmid: 4277610
references.citation: J Cell Biol. 1994 Jul;126(1):99-110
references.pmid: 8027190
references.citation: J Biol Chem. 2008 Mar 7;283(10):6393-401
references.pmid: 18165687
references.citation: Annu Rev Biochem. 2000;69:247-75
references.pmid: 10966459
references.citation: Science. 2006 Sep 15;313(5793):1636-7
references.pmid: 16973881
references.citation: J Biol Chem. 2007 Oct 19;282(42):30466-75
references.pmid: 17726018
references.citation: Mol Biol Cell. 2004 Oct;15(10):4609-21
references.pmid: 15254266
references.citation: Mol Biol Cell. 1999 Nov;10(11):3943-57
references.pmid: 10564282
references.citation: EMBO J. 2001 Aug 1;20(15):4035-40
references.pmid: 11483507
references.citation: Traffic. 2000 Aug;1(8):605-13
references.pmid: 11208148
references.citation: EMBO J. 2002 Mar 15;21(6):1241-7
references.pmid: 11889030
references.citation: Clin Microbiol Rev. 1999 Oct;12(4):501-17
references.pmid: 10515900
references.citation: Biochim Biophys Acta. 1985 Dec 4;837(3):336-43
references.pmid: 3904834
references.citation: Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11194-9
references.pmid: 10500153
references.citation: Nature. 1998 Dec 10;396(6711):575-80
references.pmid: 9859992
references.citation: J Cell Biol. 2008 Feb 25;180(4):813-26
references.pmid: 18299351
references.citation: Biochim Biophys Acta. 1976 Dec 2;455(2):452-65
references.pmid: 793632
references.citation: J Appl Microbiol. 2009 Jun;106(6):1908-18
references.pmid: 19228256
references.citation: J Microbiol Methods. 2008 Aug;74(2-3):64-73
references.pmid: 18485505
references.citation: Biochim Biophys Acta. 1974 Feb 26;339(1):57-70
references.pmid: 4546885
references.citation: Annu Rev Cell Dev Biol. 1998;14:111-36
references.pmid: 9891780
references.citation: Biochim Biophys Acta. 1986 Dec 22;864(3-4):257-304
references.pmid: 3539192
references.citation: J Cell Biol. 1988 Oct;107(4):1369-83
references.pmid: 3049619
references.citation: Mol Biol Cell. 2002 Aug;13(8):2664-80
references.pmid: 12181337
references.citation: Nature. 2001 Feb 1;409(6820):581-8
references.pmid: 11214310
references.citation: Yeast. 1998 Jan 30;14(2):115-32
references.pmid: 9483801
references.citation: Nature. 1997 Jun 5;387(6633):569-72
references.pmid: 9177342
references.citation: Annu Rev Cell Dev Biol. 2002;18:289-314
references.pmid: 12142286
references.citation: Mol Biol Cell. 2007 Oct;18(10):3873-82
references.pmid: 17652457
references.citation: Autophagy. 2008 Jan;4(1):5-19
references.pmid: 17932463
references.citation: EMBO J. 2001 Oct 15;20(20):5650-6
references.pmid: 11598008
references.citation: Traffic. 2004 Apr;5(4):255-64
references.pmid: 15030567