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The Essentials of Protein Import in the Degenerate Mitochondrion of


Several essential biochemical processes are situated in mitochondria. The metabolic transformation of mitochondria in distinct lineages of eukaryotes created proteomes ranging from thousands of proteins to what appear to be a much simpler scenario. In the case of Entamoeba histolytica, tiny mitochondria known as mitosomes have undergone extreme reduction. Only recently a single complete metabolic pathway of sulfate activation has been identified in these organelles. The E. histolytica mitosomes do not produce ATP needed for the sulfate activation pathway and for three molecular chaperones, Cpn60, Cpn10 and mtHsp70. The already characterized ADP/ATP carrier would thus be essential to provide cytosolic ATP for these processes, but how the equilibrium of inorganic phosphate could be maintained was unknown. Finally, how the mitosomal proteins are translocated to the mitosomes had remained unclear. We used a hidden Markov model (HMM) based search of the E. histolytica genome sequence to discover candidate (i) mitosomal phosphate carrier complementing the activity of the ADP/ATP carrier and (ii) membrane-located components of the protein import machinery that includes the outer membrane translocation channel Tom40 and membrane assembly protein Sam50. Using in vitro and in vivo systems we show that E. histolytica contains a minimalist set up of the core import components in order to accommodate a handful of mitosomal proteins. The anaerobic and parasitic lifestyle of E. histolytica has produced one of the simplest known mitochondrial compartments of all eukaryotes. Comparisons with mitochondria of another amoeba, Dictystelium discoideum, emphasize just how dramatic the reduction of the protein import apparatus was after the loss of archetypal mitochondrial functions in the mitosomes of E. histolytica.


Vyšlo v časopise: The Essentials of Protein Import in the Degenerate Mitochondrion of. PLoS Pathog 6(3): e32767. doi:10.1371/journal.ppat.1000812
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000812

Souhrn

Several essential biochemical processes are situated in mitochondria. The metabolic transformation of mitochondria in distinct lineages of eukaryotes created proteomes ranging from thousands of proteins to what appear to be a much simpler scenario. In the case of Entamoeba histolytica, tiny mitochondria known as mitosomes have undergone extreme reduction. Only recently a single complete metabolic pathway of sulfate activation has been identified in these organelles. The E. histolytica mitosomes do not produce ATP needed for the sulfate activation pathway and for three molecular chaperones, Cpn60, Cpn10 and mtHsp70. The already characterized ADP/ATP carrier would thus be essential to provide cytosolic ATP for these processes, but how the equilibrium of inorganic phosphate could be maintained was unknown. Finally, how the mitosomal proteins are translocated to the mitosomes had remained unclear. We used a hidden Markov model (HMM) based search of the E. histolytica genome sequence to discover candidate (i) mitosomal phosphate carrier complementing the activity of the ADP/ATP carrier and (ii) membrane-located components of the protein import machinery that includes the outer membrane translocation channel Tom40 and membrane assembly protein Sam50. Using in vitro and in vivo systems we show that E. histolytica contains a minimalist set up of the core import components in order to accommodate a handful of mitosomal proteins. The anaerobic and parasitic lifestyle of E. histolytica has produced one of the simplest known mitochondrial compartments of all eukaryotes. Comparisons with mitochondria of another amoeba, Dictystelium discoideum, emphasize just how dramatic the reduction of the protein import apparatus was after the loss of archetypal mitochondrial functions in the mitosomes of E. histolytica.


Zdroje

1. EmbleyTM

MartinW

2006 Eukaryotic evolution, changes and challenges. Nature 440 623 630

2. Cavalier-SmithT

1989 Molecular phylogeny. Archaebacteria and Archezoa. Nature 339 l00 l01

3. AkhmanovaA

VonckenF

van AlenT

vanHA

BoxmaB

1998 A hydrogenosome with a genome. Nature 396 527 528

4. HrdýI

HirtRP

DolezalP

BardonováL

FosterPG

2004 Trichomonas hydrogenosomes contain the NADH dehydrogenase module of mitochondrial complex I. Nature 432 618 622

5. DyallSD

YanW

Delgadillo-CorreaMG

LuncefordA

LooJA

2004 Non-mitochondrial complex I proteins in a hydrogenosomal oxidoreductase complex. Nature 431 1103 1107

6. van derGiezen

2009 Hydrogenosomes and mitosomes: conservation and evolution of functions. J Eukaryot Microbiol 56 221 231

7. MokranjacD

NeupertW

2005 Protein import into mitochondria. Biochem Soc Trans 33 1019 1023

8. KutikS

GuiardB

MeyerHE

WiedemannN

PfannerN

2007 Cooperation of translocase complexes in mitochondrial protein import. J Cell Biol 179 585 591

9. DolezalP

LikićV

TachezyJ

LithgowT

2006 Evolution of the molecular machines for protein import into mitochondria. Science 313 314 318

10. DolezalP

SmidO

RadaP

ZubacováZ

BursaćD

2005 Giardia mitosomes and trichomonad hydrogenosomes share a common mode of protein targeting. Proc Natl Acad Sci U S A 102 10924 10929

11. RegoesA

ZourmpanouD

León-AvilaG

van derGiezen

TovarJ

2005 Protein import, replication, and inheritance of a vestigial mitochondrion. J Biol Chem 280 30557 30563

12. BurriL

WilliamsBA

BursaćD

LithgowT

KeelingPJ

2006 Microsporidian mitosomes retain elements of the general mitochondrial targeting system. Proc Natl Acad Sci U S A 103 15916 15920

13. BurriL

KeelingPJ

2007 Protein targeting in parasites with cryptic mitochondria. Int J Parasitol 37 265 272

14. TovarJ

León-AvilaG

SanchezLB

SutakR

TachezyJ

2003 Mitochondrial remnant organelles of Giardia function in iron-sulphur protein maturation. Nature 426 172 176

15. TsaousisAD

KunjiER

GoldbergAV

LucocqJM

HirtRP

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

16. WilliamsBA

HirtRP

LucocqJM

EmbleyTM

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

17. TovarJ

FischerA

ClarkCG

1999 The mitosome, a novel organelle related to mitochondria in the amitochondrial parasite Entamoeba histolytica. Mol Microbiol 32 1013 1021

18. MaiZ

GhoshS

FrisardiM

RosenthalB

RogersR

1999 Hsp60 is targeted to a cryptic mitochondrion-derived organelle (“crypton”) in the microaerophilic protozoan parasite Entamoeba histolytica. Mol Cell Biol 19 2198 2205

19. GoldbergAV

MolikS

TsaousisAD

NeumannK

KuhnkeG

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

20. KatinkaMD

DupratS

CornillotE

MetenierG

ThomaratF

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

21. MaralikovaB

AliV

Nakada-TsukuiK

NozakiT

van derGiezen

2009 Bacterial-type oxygen detoxification and iron-sulfur cluster assembly in amoebal relict mitochondria. Cell Microbiol

22. Mi-IchiF

YousufMA

Nakada-TsukuiK

NozakiT

2009 Mitosomes in Entamoeba histolytica contain a sulfate activation pathway. Proc Natl Acad Sci U S A

23. LillR

MühlenhoffU

2006 Iron-sulfur protein biogenesis in eukaryotes: components and mechanisms. Annu Rev Cell Dev Biol 22 457 486

24. ClarkCG

2000 The evolution of Entamoeba, a cautionary tale. Res Microbiol 151 599 603

25. van der GiezenM

CoxS

TovarJ

2004 The iron-sulfur cluster assembly genes iscS and iscU of Entamoeba histolytica were acquired by horizontal gene transfer. BMC Evol Biol 4 7

26. AliV

ShigetaY

TokumotoU

TakahashiY

NozakiT

2004 An intestinal parasitic protist, Entamoeba histolytica, possesses a non-redundant nitrogen fixation-like system for iron-sulfur cluster assembly under anaerobic conditions. J Biol Chem 279 16863 16874

27. ChanKW

SlotboomDJ

CoxS

EmbleyTM

FabreO

2005 A novel ADP/ATP transporter in the mitosome of the microaerophilic human parasite Entamoeba histolytica. Curr Biol 15 737 742

28. TovarJ

CoxSS

van der GiezenM

2007 A mitosome purification protocol based on percoll density gradients and its use in validating the mitosomal nature of Entamoeba histolytica mitochondrial Hsp70. Methods Mol Biol 390 167 177

29. van der GiezenM

León-AvilaG

TovarJ

2005 Characterization of chaperonin 10 (Cpn10) from the intestinal human pathogen Entamoeba histolytica. Microbiology 151 3107 3115

30. Pebay-PeyroulaE

Dahout-GonzalezC

KahnR

TrézéguetV

LauquinGJ

2003 Structure of mitochondrial ADP/ATP carrier in complex with carboxyatractyloside. Nature 426 39 44

31. SatreM

MatteiS

AubryL

GaudetP

PelosiL

2007 Mitochondrial carrier family: repertoire and peculiarities of the cellular slime mould Dictyostelium discoideum. Biochimie 89 1058 1069

32. KunjiER

RobinsonAJ

2006 The conserved substrate binding site of mitochondrial carriers. Biochim Biophys Acta 1757 1237 1248

33. HamelP

Saint-GeorgesY

de PintoB

LachacinskiN

AltamuraN

2004 Redundancy in the function of mitochondrial phosphate transport in Saccharomyces cerevisiae and Arabidopsis thaliana. Mol Microbiol 51 307 317

34. MurakamiH

BlobelG

PainD

1990 Isolation and characterization of the gene for a yeast mitochondrial import receptor. Nature 347 488 491

35. SimpsonAG

RogerAJ

2002 Eukaryotic evolution: getting to the root of the problem. Curr Biol 12 R691 R693

36. NikolaevSI

BerneyC

PetrovNB

MylnikovAP

FahrniJF

2006 Phylogenetic position of Multicilia marina and the evolution of Amoebozoa. Int J Syst Evol Microbiol 56 1449 1458

37. EichingerL

PachebatJA

GlocknerG

RajandreamMA

SucgangR

2005 The genome of the social amoeba Dictyostelium discoideum. Nature 435 43 57

38. MacasevD

WhelanJ

NewbiginE

Silva-FilhoMC

MulhernTD

2004 Tom22′, an 8-kDa trans-site receptor in plants and protozoans, is a conserved feature of the TOM complex that appeared early in the evolution of eukaryotes. Mol Biol Evol 21 1557 1564

39. BarthC

LeP

FisherPR

2007 Mitochondrial biology and disease in Dictyostelium. Int Rev Cytol 263 207 252

40. NagayamaK

ItonoS

YoshidaT

IshiguroS

OchiaiH

2008 Antisense RNA inhibition of the beta subunit of the Dictyostelium discoideum mitochondrial processing peptidase induces the expression of mitochondrial proteins. Biosci Biotechnol Biochem 72 1836 1846

41. LoftusB

AndersonI

DaviesR

AlsmarkUC

SamuelsonJ

2005 The genome of the protist parasite Entamoeba histolytica. Nature 433 865 868

42. BakatselouC

KidgellC

GrahamCC

2000 A mitochondrial-type hsp70 gene of Entamoeba histolytica. Mol Biochem Parasitol 110 177 182

43. WallerRF

JabbourC

ChanNC

CelikN

LikićVA

2009 Evidence of a reduced and modified mitochondrial protein import apparatus in microsporidian mitosomes. Eukaryot Cell 8 19 26

44. PusnikM

CharriereF

MaserP

WallerRF

DagleyMJ

2009 The single mitochondrial porin of Trypanosoma brucei is the main metabolite transporter in the outer mitochondrial membrane. Mol Biol Evol 26 671 680

45. FinnRD

TateJ

MistryJ

CoggillPC

SammutSJ

2008 The Pfam protein families database. Nucleic Acids Res 36 D281 D288

46. FrickeyT

LupasA

2004 CLANS: a Java application for visualizing protein families based on pairwise similarity. Bioinformatics 20 3702 3704

47. DagleyMJ

DolezalP

LikićVA

SmidO

PurcellAW

2009 The protein import channel in the outer mitosomal membrane of Giardia intestinalis. Mol Biol Evol 26 1941 1947

48. KutikS

StojanovskiD

BeckerL

BeckerT

MeineckeM

2008 Dissecting membrane insertion of mitochondrial beta-barrel proteins. Cell 132 1011 1024

49. GabrielK

EganB

LithgowT

2003 Tom40, the import channel of the mitochondrial outer membrane, plays an active role in sorting imported proteins. EMBO J 22 2380 2386

50. WilihoeftU

Campos-GongoraE

TouzniS

BruchhausI

TannichE

2001 Introns of Entamoeba histolytica and Entamoeba dispar. Protist 152 149 156

51. GentleIE

BurriL

LithgowT

2005 Molecular architecture and function of the Omp85 family of proteins. Mol Microbiol 58 1216 1225

52. VoulhouxR

TommassenJ

2004 Omp85, an evolutionarily conserved bacterial protein involved in outer-membrane-protein assembly. Res Microbiol 155 129 135

53. GentleI

GabrielK

BeechP

WallerR

LithgowT

2004 The Omp85 family of proteins is essential for outer membrane biogenesis in mitochondria and bacteria. J Cell Biol 164 19 24

54. Sánchez-PulidoL

DevosD

GenevroisS

VicenteM

ValenciaA

2003 POTRA: a conserved domain in the FtsQ family and a class of beta-barrel outer membrane proteins. Trends Biochem Sci 28 523 526

55. PicazarriK

Nakada-TsukuiK

NozakiT

2008 Autophagy during proliferation and encystation in the protozoan parasite Entamoeba invadens. Infect Immun 76 278 288

56. León-AvilaG

TovarJ

2004 Mitosomes of Entamoeba histolytica are abundant mitochondrion-related remnant organelles that lack a detectable organellar genome. Microbiology 150 1245 1250

57. van derGiezen

BirdseyGM

HornerDS

LucocqJ

DyalPL

2003 Fungal hydrogenosomes contain mitochondrial heat-shock proteins. Mol Biol Evol 20 1051 1061

58. EddySR

1996 Hidden Markov models. Curr Opin Struct Biol 6 361 365

59. SchneiderA

BursaćD

LithgowT

2008 The direct route: a simplified pathway for protein import into the mitochondrion of trypanosomes. Trends Cell Biol 18 12 18

60. SmidO

MatuskovaA

HarrisSR

KuceraT

NovotnyM

2008 Reductive evolution of the mitochondrial processing peptidases of the unicellular parasites trichomonas vaginalis and Giardia intestinalis. PLoS Pathog 4 e1000243 doi:10.1371/journal.ppat.1000243

61. SaavedraE

EncaladaR

PinedaE

Jasso-ChavezR

Moreno-SánchezR

2005 Glycolysis in Entamoeba histolytica. Biochemical characterization of recombinant glycolytic enzymes and flux control analysis. FEBS J 272 1767 1783

62. ChacinskaA

KoehlerCM

MilenkovicD

LithgowT

PfannerN

2009 Importing mitochondrial proteins: machineries and mechanisms. Cell 138 628 644

63. LikićVA

PerryA

HulettJ

DerbyM

TravenA

2005 Patterns that define the four domains conserved in known and novel isoforms of the protein import receptor Tom20. J Mol Biol 347 81 93

64. ChanNC

LikićVA

WallerRF

MulhernTD

LithgowT

2006 The C-terminal TPR domain of Tom70 defines a family of mitochondrial protein import receptors found only in animals and fungi. J Mol Biol 358 1010 1022

65. GillEE

az-TrivinoS

BarberaMJ

SilbermanJD

StechmannA

2007 Novel mitochondrion-related organelles in the anaerobic amoeba Mastigamoeba balamuthi. Mol Microbiol 66 1306 1320

66. EddySR

1998 Profile hidden Markov models. Bioinformatics 14 755 763

67. ArnoldK

BordoliL

KoppJ

SchwedeT

2006 The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics 22 195 201

68. ThompsonJD

GibsonTJ

PlewniakF

JeanmouginF

HigginsDG

1997 The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25 4876 4882

69. HusonDH

BryantD

2006 Application of phylogenetic networks in evolutionary studies. Mol Biol Evol 23 254 267

70. HuelsenbeckJP

RonquistF

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

71. DiamondLS

HarlowDR

CunnickCC

1978 A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg 72 431 432

72. LithgowT

JunneT

WachterC

SchatzG

1994 Yeast mitochondria lacking the two import receptors Mas20p and Mas70p can efficiently and specifically import precursor proteins. J Biol Chem 269 15325 15330

73. DaumG

BohniPC

SchatzG

1982 Import of proteins into mitochondria. Cytochrome b2 and cytochrome c peroxidase are located in the intermembrane space of yeast mitochondria. J Biol Chem 257 13028 13033

74. BeilharzT

EganB

SilverPA

HofmannK

LithgowT

2003 Bipartite signals mediate subcellular targeting of tail-anchored membrane proteins in Saccharomyces cerevisiae. J Biol Chem 278 8219 8223

75. HamannL

NickelR

TannichE

1995 Transfection and continuous expression of heterologous genes in the protozoan parasite Entamoeba histolytica. Proc Natl Acad Sci U S A 92 8975 8979

76. NijtmansLG

HendersonNS

HoltIJ

2002 Blue Native electrophoresis to study mitochondrial and other protein complexes. Methods 26 327 334

77. WittigI

BraunHP

SchaggerH

2006 Blue native PAGE. Nat Protoc 1 418 428

78. GabrielK

PfannerN

2007 The mitochondrial machinery for import of precursor proteins. Methods Mol Biol 390 99 117

79. YoukerRT

WalshP

BeilharzT

LithgowT

BrodskyJL

2004 Distinct roles for the Hsp40 and Hsp90 molecular chaperones during cystic fibrosis transmembrane conductance regulator degradation in yeast. Mol Biol Cell 15 4787 4797

80. GuindonS

GascuelO

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

81. SodingJ

BiegertA

LupasAN

2005 The HHpred interactive server for protein homology detection and structure prediction. Nucleic Acids Res 33 W244 W248

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