title: The involvement of the Mid1/Cch1/Yvc1 calcium channels in Aspergillus fumigatus virulence.
abstract: Aspergillus fumigatus is a major opportunistic pathogen and allergen of mammals. Calcium homeostasis and signaling is essential for numerous biological processes and also influences A. fumigatus pathogenicity. The presented study characterized the function of the A. fumigatus homologues of three Saccharomyces cerevisiae calcium channels, voltage-gated Cch1, stretch-activated Mid1 and vacuolar Yvc1. The A. fumigatus calcium channels cchA, midA and yvcA were regulated at transcriptional level by increased calcium levels. The YvcA::GFP fusion protein localized to the vacuoles. Both ΔcchA and ΔmidA mutant strains showed reduced radial growth rate in nutrient-poor minimal media. Interestingly, this growth defect in the ΔcchA strain was rescued by the exogenous addition of CaCl2. The ΔcchA, ΔmidA, and ΔcchA ΔmidA strains were also sensitive to the oxidative stress inducer, paraquat. Restriction of external Ca(2+) through the addition of the Ca(2+)-chelator EGTA impacted upon the growth of the ΔcchA and ΔmidA strains. All the A. fumigatus ΔcchA, ΔmidA, and ΔyvcA strains demonstrated attenuated virulence in a neutropenic murine model of invasive pulmonary aspergillosis. Infection with the parental strain resulted in a 100% mortality rate at 15 days post-infection, while the mortality rate of the ΔcchA, ΔmidA, and ΔyvcA strains after 15 days post-infection was only 25%. Collectively, this investigation strongly indicates that CchA, MidA, and YvcA play a role in A. fumigatus calcium homeostasis and virulence.
authors.affiliation: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.
authors.forename: Patrícia Alves
authors.initials: PA
authors.lastname: de Castro
authors.affiliation: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.
authors.forename: Jéssica
authors.initials: J
authors.lastname: Chiaratto
authors.affiliation: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.
authors.forename: Lizziane K
authors.initials: LK
authors.lastname: Winkelströter
authors.affiliation: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.
authors.forename: Vinícius Leite Pedro
authors.initials: VL
authors.lastname: Bom
authors.affiliation: Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.
authors.forename: Leandra Naira Zambelli
authors.initials: LN
authors.lastname: Ramalho
authors.affiliation: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.
authors.forename: Maria Helena S
authors.initials: MH
authors.lastname: Goldman
authors.affiliation: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.
authors.forename: Neil Andrew
authors.initials: NA
authors.lastname: Brown
authors.affiliation: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil; National Laboratory of Science and Technology of Bioethanol (CTBE), Campinas, Brazil.
authors.forename: Gustavo H
authors.initials: GH
authors.lastname: Goldman
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D001232:Aspergillus fumigatus / Q000235:genetics / Q000254:growth & development / Q000378:metabolism* / Q000472:pathogenicity*; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism*; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008807:Mice, Inbred BALB C; D009154:Mutation / Q000235:genetics; D009503:Neutropenia / Q000382:microbiology / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014774:Virulence / Q000187:drug effects
chemical_list: D000935:Antifungal Agents; D015220:Calcium Channels; D005656:Fungal Proteins; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins
references.citation: BMC Genomics. 2014 Feb 25;15:159
references.pmid: 24568630
references.citation: Microbiology (Reading). 2009 Dec;155(Pt 12):3922-3933
references.pmid: 19762439
references.citation: PLoS Pathog. 2013;9(8):e1003564
references.pmid: 23990785
references.citation: J Cell Sci. 2013 May 15;126(Pt 10):2141-50
references.pmid: 23729741
references.citation: Eukaryot Cell. 2008 Feb;7(2):415-24
references.pmid: 18083828
references.citation: Eukaryot Cell. 2011 Jun;10(6):832-41
references.pmid: 21357477
references.citation: FEBS Lett. 2005 Oct 24;579(25):5697-703
references.pmid: 16223494
references.citation: J Med Microbiol. 2007 Sep;56(Pt 9):1205-1212
references.pmid: 17761484
references.citation: Mol Cell Biol. 2000 Sep;20(18):6686-94
references.pmid: 10958666
references.citation: FEMS Yeast Res. 2011 Aug;11(5):430-9
references.pmid: 21457451
references.citation: Eukaryot Cell. 2008 Jul;7(7):1085-97
references.pmid: 18456861
references.citation: Mycopathologia. 2012 Dec;174(5-6):359-69
references.pmid: 22886468
references.citation: Fungal Genet Biol. 2007 Jun;44(6):554-62
references.pmid: 17118679
references.citation: PLoS One. 2012;7(10):e46564
references.pmid: 23071589
references.citation: J Biol Chem. 2010 Apr 2;285(14):10951-8
references.pmid: 20123986
references.citation: Fungal Genet Biol. 2009 Oct;46(10):791-802
references.pmid: 19573616
references.citation: FEBS Lett. 1997 Dec 15;419(2-3):259-62
references.pmid: 9428646
references.citation: J Allergy Clin Immunol. 2002 Nov;110(5):685-92
references.pmid: 12417875
references.citation: Eukaryot Cell. 2008 May;7(5):747-64
references.pmid: 18375617
references.citation: Mol Microbiol. 2006 Mar;59(5):1429-51
references.pmid: 16468987
references.citation: Cell Calcium. 2011 Jun;49(6):376-86
references.pmid: 21511333
references.citation: Front Microbiol. 2013 Jun 06;4:141
references.pmid: 23760756
references.citation: Int J Med Microbiol. 2014 May;304(3-4):339-50
references.pmid: 24368068
references.citation: Commun Integr Biol. 2011 Mar;4(2):186-7
references.pmid: 21655435
references.citation: Mol Microbiol. 2008 Mar;67(6):1274-91
references.pmid: 18298443
references.citation: Exp Cell Res. 2005 Nov 15;311(1):84-95
references.pmid: 16202999
references.citation: Mol Microbiol. 1990 May;4(5):839-43
references.pmid: 2388561
references.citation: Eukaryot Cell. 2006 Oct;5(10):1788-96
references.pmid: 16950930
references.citation: J Biol Chem. 2003 Oct 3;278(40):38461-9
references.pmid: 12878605
references.citation: Cell Calcium. 2011 Aug;50(2):139-46
references.pmid: 21310481
references.citation: J Biol Chem. 2011 Mar 25;286(12):10744-54
references.pmid: 21252230
references.citation: Curr Genet. 1989 Dec;16(5-6):339-46
references.pmid: 2692852
references.citation: Bioessays. 2002 Oct;24(10):894-903
references.pmid: 12325122
references.citation: Eukaryot Cell. 2013 May;12(5):712-24
references.pmid: 23475703
references.citation: Biochem Biophys Res Commun. 2003 Nov 28;311(4):1143-50
references.pmid: 14623300
references.citation: Nat Rev Mol Cell Biol. 2000 Oct;1(1):11-21
references.pmid: 11413485
references.citation: Fungal Genet Biol. 2009 Apr;46(4):287-98
references.pmid: 19570501
references.citation: PLoS One. 2012;7(5):e37591
references.pmid: 22649543
references.citation: Mol Genet Genomics. 2008 Dec;280(6):483-95
references.pmid: 18830711
references.citation: Antioxid Redox Signal. 2014 Jul 1;21(1):123-37
references.pmid: 24512092
references.citation: Mol Microbiol. 2008 Jun;68(6):1348-65
references.pmid: 18433453
references.citation: Proc Natl Acad Sci U S A. 2006 Jul 5;103(27):10352-10357
references.pmid: 16801547
references.citation: Mol Cell Biol. 1997 Nov;17(11):6339-47
references.pmid: 9343395
references.citation: Adv Genet. 1977;19:33-131
references.pmid: 327767
references.citation: Genes Dev. 1999 Apr 1;13(7):798-803
references.pmid: 10197980
references.citation: FEBS Lett. 2004 Oct 22;576(3):291-6
references.pmid: 15498550
references.citation: Fungal Genet Biol. 2013 Aug;57:23-32
references.pmid: 23769872
references.citation: Biochem Biophys Res Commun. 2003 Nov 28;311(4):1151-7
references.pmid: 14623301
references.citation: Eukaryot Cell. 2010 Mar;9(3):472-6
references.pmid: 20097742
references.citation: Eukaryot Cell. 2008 Apr;7(4):647-55
references.pmid: 18296620
references.citation: Front Biosci (Landmark Ed). 2012 Jan 01;17:1079-93
references.pmid: 22201791
references.citation: J Cell Biol. 2002 Jan 7;156(1):29-34
references.pmid: 11781332
references.citation: FEBS Lett. 2010 May 17;584(10):2028-32
references.pmid: 20035756
references.citation: Fungal Genet Biol. 2004 Sep;41(9):827-41
references.pmid: 15288019
references.citation: J Biol Chem. 2002 Aug 23;277(34):31079-88
references.pmid: 12058033
references.citation: Mol Microbiol. 2011 Dec;82(5):1235-59
references.pmid: 22066998
references.citation: Mol Cell Biol. 1994 Dec;14(12):8259-71
references.pmid: 7526155
references.citation: FEMS Microbiol Lett. 2007 Jul;272(1):1-7
references.pmid: 17517068
references.citation: Nat Rev Mol Cell Biol. 2003 Jul;4(7):517-29
references.pmid: 12838335
references.citation: BMC Microbiol. 2010 Jan 15;10:12
references.pmid: 20078882
references.citation: Nat Rev Microbiol. 2007 Jun;5(6):418-30
references.pmid: 17505522
references.citation: Fungal Genet Biol. 2007 Mar;44(3):219-30
references.pmid: 16990036
references.citation: Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):7105-10
references.pmid: 12771382
references.citation: Proc Natl Acad Sci U S A. 2001 Jul 3;98(14):7801-5
references.pmid: 11427713
references.citation: Microbiology (Reading). 2008 Dec;154(Pt 12):3775-3781
references.pmid: 19047745
references.citation: Fungal Genet Biol. 2014 Jan;62:62-70
references.pmid: 24239700
references.citation: Clin Microbiol Rev. 2009 Jul;22(3):447-65
references.pmid: 19597008
references.citation: Eukaryot Cell. 2003 Feb;2(1):34-48
references.pmid: 12582121
references.citation: Genetics. 2001 Dec;159(4):1527-38
references.pmid: 11779794
references.citation: Eukaryot Cell. 2006 Feb;5(2):411-21
references.pmid: 16467481
references.citation: Appl Environ Microbiol. 2002 Mar;68(3):1351-7
references.pmid: 11872487
references.citation: FEMS Yeast Res. 2014 Jun;14(4):633-41
references.pmid: 24650198
references.citation: J Biol Chem. 2004 Oct 15;279(42):43614-24
references.pmid: 15299026
references.citation: Eukaryot Cell. 2012 Aug;11(8):978-88
references.pmid: 22635922