A Broad Distribution of the Alternative Oxidase in Microsporidian Parasites

English version České info

Microsporidia are a group of obligate intracellular parasitic eukaryotes that were considered to be amitochondriate until the recent discovery of highly reduced mitochondrial organelles called mitosomes. Analysis of the complete genome of Encephalitozoon cuniculi revealed a highly reduced set of proteins in the organelle, mostly related to the assembly of iron-sulphur clusters. Oxidative phosphorylation and the Krebs cycle proteins were absent, in keeping with the notion that the microsporidia and their mitosomes are anaerobic, as is the case for other mitosome bearing eukaryotes, such as Giardia. Here we provide evidence opening the possibility that mitosomes in a number of microsporidian lineages are not completely anaerobic. Specifically, we have identified and characterized a gene encoding the alternative oxidase (AOX), a typically mitochondrial terminal oxidase in eukaryotes, in the genomes of several distantly related microsporidian species, even though this gene is absent from the complete genome of E. cuniculi. In order to confirm that these genes encode functional proteins, AOX genes from both A. locustae and T. hominis were over-expressed in E. coli and AOX activity measured spectrophotometrically using ubiquinol-1 (UQ-1) as substrate. Both A. locustae and T. hominis AOX proteins reduced UQ-1 in a cyanide and antimycin-resistant manner that was sensitive to ascofuranone, a potent inhibitor of the trypanosomal AOX. The physiological role of AOX microsporidia may be to reoxidise reducing equivalents produced by glycolysis, in a manner comparable to that observed in trypanosomes.

Vyšlo v časopise: A Broad Distribution of the Alternative Oxidase in Microsporidian Parasites. PLoS Pathog 6(2): e32767. doi:10.1371/journal.ppat.1000761
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000761

Souhrn

Zdroje

1. DidierES

WeissLM

2006 Microsporidiosis: current status. Curr Opin Infect Dis 19 485 492

2. KeelingPJ

DoolittleWF

1996 Alpha-tubulin from early-diverging eukaryotic lineages and the evolution of the tubulin family. Mol Biol Evol 13 1297 1305

3. HirtRP

LogsdonJMJr

HealyB

DoreyMW

DoolittleWF

1999 Microsporidia are related to Fungi: evidence from the largest subunit of RNA polymerase II and other proteins. Proc Natl Acad Sci U S A 96 580 585

4. EdlindTD

LiJ

VisvesvaraGS

VodkinMH

McLaughlinGL

1996 Phylogenetic analysis of beta-tubulin sequences from amitochondrial protozoa. Mol Phylogenet Evol 5 359 367

5. ThomaratF

VivaresCP

GouyM

2004 Phylogenetic analysis of the complete genome sequence of Encephalitozoon cuniculi supports the fungal origin of microsporidia and reveals a high frequency of fast-evolving genes. J Mol Evol 59 780 791

6. KatinkaMD

DupratS

CornillotE

MetenierG

ThomaratF

2001 Genome sequence and gene compaction of the eukaryote parasite Encephalitozoon cuniculi. Nature 414 450 453

7. BeznoussenkoGV

DolgikhVV

SeliverstovaEV

SemenovPB

TokarevYS

2007 Analogs of the Golgi complex in microsporidia: structure and avesicular mechanisms of function. J Cell Sci 120 1288 1298

8. WilliamsBA

HirtRP

LucocqJM

EmbleyTM

2002 A mitochondrial remnant in the microsporidian Trachipleistophora hominis. Nature 418 865 869

9. WeidnerE

FindleyAM

DolgikhV

SokolovaJ

1999 Microsporidian biochemistry and physiology.

WittnerM

WeissLM

The Microsporidia and Microsporidiosis Washington, D.C. ASM Press

10. TsaousisAD

KunjiER

GoldbergAV

LucocqJM

HirtRP

2008 A novel route for ATP acquisition by the remnant mitochondria of Encephalitozoon cuniculi. Nature 453 553 556

11. RichardsTA

HirtRP

WilliamsBA

EmbleyTM

2003 Horizontal gene transfer and the evolution of parasitic protozoa. Protist 154 17 32

12. WilliamsBA

CaliA

TakvorianPM

KeelingPJ

2008 Distinct localization patterns of two putative mitochondrial proteins in the microsporidian Encephalitozoon cuniculi. J Eukaryot Microbiol 55 131 133

13. GoldbergAV

MolikS

TsaousisAD

NeumannK

KuhnkeG

2008 Localization and functionality of microsporidian iron-sulphur cluster assembly proteins. Nature 452 624 628

14. LarssonC

PahlmanIL

AnsellR

RigouletM

AdlerL

1998 The importance of the glycerol 3-phosphate shuttle during aerobic growth of Saccharomyces cerevisiae. Yeast 14 347 357

15. SlamovitsCH

FastNM

LawJS

KeelingPJ

2004 Genome compaction and stability in microsporidian intracellular parasites. Curr Biol 14 891 896

16. ChaudhuriM

OttRD

HillGC

2006 Trypanosome alternative oxidase: from molecule to function. Trends Parasitol 22 484 491

17. SuzukiT

HashimotoT

YabuY

KidoY

SakamotoK

2004 Direct evidence for cyanide-insensitive quinol oxidase (alternative oxidase) in apicomplexan parasite Cryptosporidium parvum: phylogenetic and therapeutic implications. Biochem Biophys Res Commun 313 1044 1052

18. AffourtitC

AlburyMS

CrichtonPG

MooreAL

2002 Exploring the molecular nature of alternative oxidase regulation and catalysis. FEBS Lett 510 121 126

19. MooreAL

AlburyMS

2008 Further insights into the structure of the alternative oxidase: from plants to parasites. Biochem Soc Trans 36 1022 1026

20. AlburyMS

ElliottC

MooreAL

2009 Towards a structural elucidation of the alternative oxidase in plants. Physiol Plant

21. StenmarkP

NordlundP

2003 A prokaryotic alternative oxidase present in the bacterium Novosphingobium aromaticivorans. FEBS Lett 552 189 192

22. VanlerbergheGC

McIntoshL

1997 Alternative oxidase: From gene to function. Annu Rev Plant Phys 48 703 734

23. JarmuszkiewiczW

WagnerAM

WagnerMJ

HryniewieckaL

1997 Immunological identification of the alternative oxidase of Acanthamoeba castellanii mitochondria. FEBS Lett 411 110 114

24. StechmannA

HamblinK

Perez-BrocalV

GastonD

RichmondGS

2008 Organelles in Blastocystis that blur the distinction between mitochondria and hydrogenosomes. Curr Biol 18 580 585

25. McDonaldAE

VanlerbergheGC

StaplesJF

2009 Alternative oxidase in animals: unique characteristics and taxonomic distribution. J Exp Biol 212 2627 2634

26. GardnerMJ

HallN

FungE

WhiteO

BerrimanM

2002 Genome sequence of the human malaria parasite Plasmodium falciparum. Nature 419 498 511

27. RobertsCW

RobertsF

HenriquezFL

AkiyoshiD

SamuelBU

2004 Evidence for mitochondrial-derived alternative oxidase in the apicomplexan parasite Cryptosporidium parvum: a potential anti-microbial agent target. Int J Parasitol 34 297 308

28. EisenJA

CoyneRS

WuM

WuD

ThiagarajanM

2006 Macronuclear genome sequence of the ciliate Tetrahymena thermophila, a model eukaryote. PLoS Biol 4 e286 doi:10.1371/journal.pbio.0040286

29. AtteiaA

van LisR

van HellemondJJ

TielensAGM

MartinW

2004 Identification of prokaryotic homologues indicates an endosymbiotic origin for the alternative oxidases of mitochondria (AOX) and chloroplasts (PTOX). Gene 330 143 148

30. Joseph-HorneT

HollomonDW

WoodPM

2001 Fungal respiration: a fusion of standard and alternative components. Biochim Biophys Acta 1504 179 195

31. AkiyoshiDE

MorrisonHG

LeiS

FengX

ZhangQ

2009 Genomic survey of the non-cultivatable opportunistic human pathogen, Enterocytozoon bieneusi. PLoS Pathog 5 e1000261 doi:10.1371/journal.ppat.1000261

32. CornmanRS

ChenYP

SchatzMC

StreetC

ZhaoY

2009 Genomic analyses of the microsporidian Nosema ceranae, an emergent pathogen of honey bees. PLoS Pathog 5 e1000466 doi:10.1371/journal.ppat.1000466

33. NiheiC

FukaiY

KitaK

2002 Trypanosome alternative oxidase as a target of chemotherapy. Biochim Biophys Acta 1587 234 239

34. KitaK

NiheiC

TomitsukaE

2003 Parasite mitochondria as drug target: Diversity and dynamic changes during the life cycle. Curr Med Chem 10 2535 2548

35. DidierES

1997 Effects of albendazole, fumagillin, and TNP-470 on microsporidial replication in vitro. Antimicrob Agents Chemother 41 1541 1546

36. DidierPJ

PhillipsJN

KueblerDJ

NasrM

BrindleyPJ

2006 Antimicrosporidial activities of fumagillin, TNP-470, ovalicin, and ovalicin derivatives in vitro and in vivo. Antimicrob Agents Chemother 50 2146 2155

37. MinagawaN

YabuY

KitaK

NagaiK

OhtaN

1997 An antibiotic, ascofuranone, specifically inhibits respiration and in vitro growth of long slender bloodstream forms of Trypanosoma brucei brucei. Mol Biochem Parasitol 84 271 280

38. QureshiSJ

PorteousDJ

BrookesAJ

1994 Alu-based vectorettes and splinkerettes. More efficient and comprehensive polymerase chain reaction amplification of human DNA from complex sources. Genet Anal Tech Appl 11 95 101

39. MooreAL

CarreJE

AffourtitC

AlburyMS

CrichtonPG

2008 Compelling EPR evidence that the alternative oxidase is a diiron carboxylate protein. Biochim Biophys Acta 1777 327 330

40. UmbachAL

NgVS

SiedowJN

2006 Regulation of plant alternative oxidase activity: a tale of two cysteines. Biochim Biophys Acta 1757 135 142

41. VossbrinckCR

Debrunner-VossbrinckBA

2005 Molecular phylogeny of the Microsporidia: ecological, ultrastructural and taxonomic considerations. Folia Parasitol (Praha) 52 131 142

42. FukaiY

NiheiC

KawaiK

YabuY

SuzukiT

2003 Overproduction of highly active trypanosome alternative oxidase in Escherichia coli heme-deficient mutant. Parasitology International 52 237 241

43. WilliamsBA

LeeRC

BecnelJJ

WeissLM

FastNM

2008 Genome sequence surveys of Brachiola algerae and Edhazardia aedis reveal microsporidia with low gene densities. BMC Genomics 9 200

44. AlburyMS

AffourtitC

CrichtonPG

MooreAL

2002 Structure of the plant alternative oxidase. Site-directed mutagenesis provides new information on the active site and membrane topology. J Biol Chem 277 1190 1194

45. NinaM

BernecheS

RouxB

2000 Anchoring of a monotopic membrane protein: the binding of prostaglandin H2 synthase-1 to the surface of a phospholipid bilayer. Eur Biophys J 29 439 454

46. ClarosMG

1995 MitoProt, a Macintosh application for studying mitochondrial proteins. Comput Appl Biosci 11 441 447

47. SmallI

PeetersN

LegeaiF

LurinC

2004 Predotar: A tool for rapidly screening proteomes for N-terminal targeting sequences. Proteomics 4 1581 1590

48. EmanuelssonO

NielsenH

BrunakS

von HeijneG

2000 Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. J Mol Biol 300 1005 1016

49. GeorgeR

BeddoeT

LandlK

LithgowT

1998 The yeast nascent polypeptide-associated complex initiates protein targeting to mitochondria in vivo. Proc Natl Acad Sci U S A 95 2296 2301

50. ElthonTE

NickelsRL

McIntoshL

1989 Monoclonal antibodies to the alternative oxidase of higher plant mitochondria. Plant Physiol 89 1311 1317

51. AlburyMS

DudleyP

WattsFZ

MooreAL

1996 Targeting the plant alternative oxidase protein to Schizosaccharomyces pombe mitochondria confers cyanide-insensitive respiration. J Biol Chem 271 17062 17066

52. BertholdDA

AnderssonME

NordlundP

2000 New insight into the structure and function of the alternative oxidase. Biochim Biophys Acta 1460 241 254

53. CrichtonPG

AffourtitC

AlburyMS

ElliotC

WeiDM

2009 Mutagenesis of the plant alternative oxidase reveals features important for oxygen binding and catalysis, and traps a stable ubisemiquinone in the ubiquinone binding site. Biochim Biophys Acta: In press

54. ThompsonJD

HigginsDG

GibsonTJ

1994 CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22 4673 4680

55. KeaneTM

CreeveyCJ

PentonyMM

NaughtonTJ

McInerneyJO

2006 Assessment of methods for amino acid matrix selection and their use on empirical data shows that ad hoc assumptions for choice of matrix are not justified. BMC Evol Biol 6 29

56. HuelsenbeckJP

RonquistF

2001 MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics 17 754 755

57. GuindonS

GascuelO

2003 A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 52 696 704