title: Mitochondrial function in the yeast form of the pathogenic fungus Paracoccidioides brasiliensis.
abstract: Differences between the respiratory chain of the fungus Paracoccidioides brasiliensis and its mammalian host are reported. Respiration, membrane potential, and oxidative phosphorylation in mitochondria from P. brasiliensis spheroplasts were evaluated in situ, and the presence of a complete (Complex I-V) functional respiratory chain was demonstrated. In succinate-energized mitochondria, ADP induced a transition from resting to phosphorylating respiration. The presence of an alternative NADH-ubiquinone oxidoreductase was indicated by: (i) the ability to oxidize exogenous NADH and (ii) the lack of sensitivity to rotenone and presence of sensitivity to flavone. Malate/NAD(+)-supported respiration suggested the presence of either a mitochondrial pyridine transporter or a glyoxylate pathway contributing to NADH and/or succinate production. Partial sensitivity of NADH/succinate-supported respiration to antimycin A and cyanide, as well as sensitivity to benzohydroxamic acids, suggested the presence of an alternative oxidase in the yeast form of the fungus. An increase in activity and gene expression of the alternative NADH dehydrogenase throughout the yeast's exponential growth phase was observed. This increase was coupled with a decrease in Complex I activity and gene expression of its subunit 6. These results support the existence of alternative respiratory chain pathways in addition to Complex I, as well as the utilization of NADH-linked substrates by P. brasiliensis. These specific components of the respiratory chain could be useful for further research and development of pharmacological agents against the fungus.
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: Vicente P
authors.initials: VP
authors.lastname: Martins
authors.forename: Frederico M
authors.initials: FM
authors.lastname: Soriani
authors.forename: Taisa
authors.initials: T
authors.lastname: Magnani
authors.forename: Valéria G
authors.initials: VG
authors.lastname: Tudella
authors.forename: Gustavo H
authors.initials: GH
authors.lastname: Goldman
authors.forename: Carlos
authors.initials: C
authors.lastname: Curti
authors.forename: Sérgio A
authors.initials: SA
authors.lastname: Uyemura
mesh_terms: D001203:Ascomycota / Q000145:classification* / Q000502:physiology* / Q000648:ultrastructure; D019069:Cell Respiration / Q000502:physiology; D053078:Membrane Potential, Mitochondrial / Q000502:physiology*; D008928:Mitochondria / Q000502:physiology*; D010100:Oxygen / Q000378:metabolism*; D013045:Species Specificity
chemical_list: D010100:Oxygen
references.citation: Eur J Biochem. 1986 Sep 15;159(3):519-24
references.pmid: 3758075
references.citation: J Med Vet Mycol. 1987 Oct;25(5):291-300
references.pmid: 3430291
references.citation: J Bacteriol. 1987 Sep;169(9):4055-60
references.pmid: 3624202
references.citation: Biochim Biophys Acta. 2002 Sep 10;1555(1-3):187-91
references.pmid: 12206913
references.citation: Med Mycol. 2001 Jun;39(3):233-41
references.pmid: 11446526
references.citation: J Cell Sci. 2001 Nov;114(Pt 21):3915-21
references.pmid: 11719558
references.citation: Biochimie. 2004 Jan;86(1):39-46
references.pmid: 14987799
references.citation: J Int Fed Clin Chem. 1998 Mar;10(1):24-7
references.pmid: 10181012
references.citation: FEBS Lett. 2004 Oct 8;576(1-2):151-5
references.pmid: 15474028
references.citation: J Biol Chem. 2006 Jan 20;281(3):1524-31
references.pmid: 16291748
references.citation: J Biol Chem. 1995 Dec 15;270(50):29836-42
references.pmid: 8530379
references.citation: Eukaryot Cell. 2003 Feb;2(1):34-48
references.pmid: 12582121
references.citation: Clin Microbiol Rev. 1993 Apr;6(2):89-117
references.pmid: 8472249
references.citation: J Gerontol. 1956 Jul;11(3):298-300
references.pmid: 13332224
references.citation: Biochim Biophys Acta. 2000 Aug 15;1459(2-3):274-83
references.pmid: 11004440
references.citation: J Biol Chem. 2002 Nov 8;277(45):42802-7
references.pmid: 12207028
references.citation: Proc Natl Acad Sci U S A. 1981 Jul;78(7):4596-600
references.pmid: 6945601
references.citation: Q Rev Biophys. 1992 Aug;25(3):253-324
references.pmid: 1470679
references.citation: Yeast. 2004 Jan 30;21(2):173-82
references.pmid: 14755642
references.citation: J Eukaryot Microbiol. 2005 Jul-Aug;52(4):374-81
references.pmid: 16014016
references.citation: Arch Biochem Biophys. 2004 May 1;425(1):14-24
references.pmid: 15081889
references.citation: J Biol Chem. 2004 Jan 2;279(1):385-93
references.pmid: 14561763
references.citation: FEMS Microbiol Lett. 1996 Aug 1;141(2-3):251-4
references.pmid: 8768530
references.citation: Proc Natl Acad Sci U S A. 1981 Nov;78(11):7124-8
references.pmid: 6947277
references.citation: Biochim Biophys Acta. 2000 Nov 20;1460(2-3):241-54
references.pmid: 11106766
references.citation: FEBS Lett. 2001 Nov 16;508(2):231-5
references.pmid: 11718721
references.citation: FEBS Lett. 2001 Jun 29;500(1-2):80-4
references.pmid: 11434931
references.citation: Diagn Microbiol Infect Dis. 2002 Dec;44(4):383-6
references.pmid: 12543545
references.citation: Med Mycol. 2002 Jun;40(3):225-42
references.pmid: 12146752
references.citation: Biochem J. 2001 Jun 1;356(Pt 2):595-604
references.pmid: 11368790
references.citation: J Biol Chem. 2001 Feb 9;276(6):3947-51
references.pmid: 11073955
references.citation: Int J Biochem Cell Biol. 2005 Dec;37(12):2478-503
references.pmid: 16103002
references.citation: Yeast. 2003 Apr 15;20(5):381-8
references.pmid: 12673621
references.citation: J Cell Sci. 1999 Jul;112 ( Pt 14):2347-54
references.pmid: 10381390
references.citation: J Mol Biol. 2000 Sep 15;302(2):455-64
references.pmid: 10970745
references.citation: Genetics. 1996 Jan;142(1):129-40
references.pmid: 8770590
references.citation: FEBS Lett. 1992 Aug 3;307(3):267-71
references.pmid: 1644182
references.citation: Biosci Rep. 2005 Jun-Aug;25(3-4):271-86
references.pmid: 16283557
references.citation: Adv Enzymol Relat Subj Biochem. 1956;17:65-134
references.pmid: 13313307
references.citation: Annu Rev Plant Physiol Plant Mol Biol. 1997 Jun;48:703-734
references.pmid: 15012279
references.citation: J Med Vet Mycol. 1994;32(5):381-8
references.pmid: 7844703
references.citation: FEBS Lett. 2002 Dec 18;532(3):459-64
references.pmid: 12482613
references.citation: Int J Biochem Cell Biol. 2004 Jan;36(1):162-72
references.pmid: 14592541
references.citation: Sabouraudia. 1985 Oct;23(5):323-34
references.pmid: 3906945
references.citation: Biochim Biophys Acta. 2002 Oct 3;1556(1):73-80
references.pmid: 12351220
references.citation: FEMS Yeast Res. 2003 May;3(3):239-45
references.pmid: 12689632
references.citation: Biochim Biophys Acta. 2001 Apr 2;1504(2-3):179-95
references.pmid: 11245784
references.citation: Eukaryot Cell. 2002 Oct;1(5):657-62
references.pmid: 12455685
references.citation: Biochim Biophys Acta. 2000 Jul 20;1459(1):77-87
references.pmid: 10924901
references.citation: Plant Cell. 1995 Jul;7(7):821-831
references.pmid: 12242388
references.citation: Infect Immun. 1989 Aug;57(8):2289-94
references.pmid: 2744848
references.citation: FEBS Lett. 1976 Oct 1;68(2):191-7
references.pmid: 976474
references.citation: Biochim Biophys Acta. 1969 Feb 25;172(2):331-3
references.pmid: 4304726
references.citation: Mol Genet Genomics. 2004 Jul;271(6):667-77
references.pmid: 15138890