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The Enteropathogenic Effector EspF Targets and Disrupts the Nucleolus by a Process Regulated by Mitochondrial Dysfunction


The nucleolus is a multifunctional structure within the nucleus of eukaryotic cells and is the primary site of ribosome biogenesis. Almost all viruses target and disrupt the nucleolus—a feature exclusive to this pathogen group. Here, using a combination of bio-imaging, genetic and biochemical analyses, we demonstrate that the enteropathogenic E. coli (EPEC) effector protein EspF specifically targets the nucleolus and disrupts a subset of nucleolar factors. Driven by a defined N-terminal nucleolar targeting domain, EspF causes the complete loss from the nucleolus of nucleolin, the most abundant nucleolar protein. We also show that other bacterial species disrupt the nucleolus, dependent on their ability to deliver effector proteins into the host cell. Moreover, we uncover a novel regulatory mechanism whereby nucleolar targeting by EspF is strictly controlled by EPEC's manipulation of host mitochondria. Collectively, this work reveals that the nucleolus may be a common feature of bacterial pathogenesis and demonstrates that a bacterial pathogen has evolved a highly sophisticated mechanism to enable spatio-temporal control over its virulence proteins.


Vyšlo v časopise: The Enteropathogenic Effector EspF Targets and Disrupts the Nucleolus by a Process Regulated by Mitochondrial Dysfunction. PLoS Pathog 6(6): e32767. doi:10.1371/journal.ppat.1000961
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000961

Souhrn

The nucleolus is a multifunctional structure within the nucleus of eukaryotic cells and is the primary site of ribosome biogenesis. Almost all viruses target and disrupt the nucleolus—a feature exclusive to this pathogen group. Here, using a combination of bio-imaging, genetic and biochemical analyses, we demonstrate that the enteropathogenic E. coli (EPEC) effector protein EspF specifically targets the nucleolus and disrupts a subset of nucleolar factors. Driven by a defined N-terminal nucleolar targeting domain, EspF causes the complete loss from the nucleolus of nucleolin, the most abundant nucleolar protein. We also show that other bacterial species disrupt the nucleolus, dependent on their ability to deliver effector proteins into the host cell. Moreover, we uncover a novel regulatory mechanism whereby nucleolar targeting by EspF is strictly controlled by EPEC's manipulation of host mitochondria. Collectively, this work reveals that the nucleolus may be a common feature of bacterial pathogenesis and demonstrates that a bacterial pathogen has evolved a highly sophisticated mechanism to enable spatio-temporal control over its virulence proteins.


Zdroje

1. HiscoxJA

2007 RNA viruses: hijacking the dynamic nucleolus. Nat Rev Microbiol 5 119 127

2. AhmadY

BoisvertFM

GregorP

CobleyA

LamondAI

2009 NOPdb: Nucleolar Proteome Database–2008 update. Nucleic Acids Res 37 D181 184

3. BoisvertFM

van KoningsbruggenS

NavascuesJ

LamondAI

2007 The multifunctional nucleolus. Nat Rev Mol Cell Biol 8 574 585

4. GinistyH

SicardH

RogerB

BouvetP

1999 Structure and functions of nucleolin. J Cell Sci 112 (Pt 6) 761 772

5. GalanJE

Wolf-WatzH

2006 Protein delivery into eukaryotic cells by type III secretion machines. Nature 444 567 573

6. KennyB

ValdiviaR

2009 Host-microbe interactions: bacteria. Curr Opin Microbiol 12 1 3

7. DeanP

KennyB

2009 The effector repertoire of enteropathogenic E. coli: ganging up on the host cell. Curr Opin Microbiol 12 101 109

8. ChenHD

FrankelG

2005 Enteropathogenic Escherichia coli: unravelling pathogenesis. FEMS Microbiol Rev 29 83 98

9. KennyB

DeVinneyR

SteinM

ReinscheidDJ

FreyEA

1997 Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence into mammalian cells. Cell 91 511 520

10. NougayredeJP

DonnenbergMS

2004 Enteropathogenic Escherichia coli EspF is targeted to mitochondria and is required to initiate the mitochondrial death pathway. Cell Microbiol 6 1097 1111

11. KennyB

JepsonM

2000 Targeting of an enteropathogenic Escherichia coli (EPEC) effector protein to host mitochondria. Cell Microbiol 2 579 590

12. McNamaraBP

KoutsourisA

O'ConnellCB

NougayredeJP

DonnenbergMS

2001 Translocated EspF protein from enteropathogenic Escherichia coli disrupts host intestinal barrier function. J Clin Invest 107 621 629

13. DeanP

KennyB

2004 Intestinal barrier dysfunction by enteropathogenic Escherichia coli is mediated by two effector molecules and a bacterial surface protein. Mol Microbiol 54 665 675

14. DeanP

MarescaM

SchullerS

PhillipsAD

KennyB

2006 Potent diarrheagenic mechanism mediated by the cooperative action of three enteropathogenic Escherichia coli-injected effector proteins. Proc Natl Acad Sci U S A 103 1876 1881

15. QuitardS

DeanP

MarescaM

KennyB

2006 The enteropathogenic Escherichia coli EspF effector molecule inhibits PI-3 kinase-mediated uptake independently of mitochondrial targeting. Cell Microbiol 8 972 981

16. NagaiT

AbeA

SasakawaC

2005 Targeting of enteropathogenic Escherichia coli EspF to host mitochondria is essential for bacterial pathogenesis: critical role of the 16th leucine residue in EspF. J Biol Chem 280 2998 3011

17. Ruchaud-SparaganoMH

MarescaM

KennyB

2007 Enteropathogenic Escherichia coli (EPEC) inactivate innate immune responses prior to compromising epithelial barrier function. Cell Microbiol 9 1909 1921

18. DragonF

GallagherJE

Compagnone-PostPA

MitchellBM

PorwancherKA

2002 A large nucleolar U3 ribonucleoprotein required for 18S ribosomal RNA biogenesis. Nature 417 967 970

19. PeculisBA

SteitzJA

1993 Disruption of U8 nucleolar snRNA inhibits 5.8S and 28S rRNA processing in the Xenopus oocyte. Cell 73 1233 1245

20. GrannemanS

VogelzangsJ

LuhrmannR

van VenrooijWJ

PruijnGJ

2004 Role of pre-rRNA base pairing and 80S complex formation in subnucleolar localization of the U3 snoRNP. Mol Cell Biol 24 8600 8610

21. ChenD

HuangS

2001 Nucleolar components involved in ribosome biogenesis cycle between the nucleolus and nucleoplasm in interphase cells. J Cell Biol 153 169 176

22. Steele-MortimerO

2008 The Salmonella-containing vacuole: moving with the times. Curr Opin Microbiol 11 38 45

23. HenselM

SheaJE

WatermanSR

MundyR

NikolausT

1998 Genes encoding putative effector proteins of the type III secretion system of Salmonella pathogenicity island 2 are required for bacterial virulence and proliferation in macrophages. Mol Microbiol 30 163 174

24. KuboriT

GalanJE

2003 Temporal regulation of salmonella virulence effector function by proteasome-dependent protein degradation. Cell 115 333 342

25. PatelJC

HuefferK

LamTT

GalanJE

2009 Diversification of a Salmonella virulence protein function by ubiquitin-dependent differential localization. Cell 137 283 294

26. JurisSJ

RudolphAE

HuddlerD

OrthK

DixonJE

2000 A distinctive role for the Yersinia protein kinase: actin binding, kinase activation, and cytoskeleton disruption. Proc Natl Acad Sci U S A 97 9431 9436

27. Kozjak-PavlovicV

RossK

RudelT

2008 Import of bacterial pathogenicity factors into mitochondria. Curr Opin Microbiol 11 9 14

28. SinclairJF

O'BrienAD

2002 Cell surface-localized nucleolin is a eukaryotic receptor for the adhesin intimin-gamma of enterohemorrhagic Escherichia coli O157:H7. J Biol Chem 277 2876 2885

29. ViswanathanVK

WeflenA

KoutsourisA

RoxasJL

HechtG

2008 Enteropathogenic E. coli-induced barrier function alteration is not a consequence of host cell apoptosis. Am J Physiol Gastrointest Liver Physiol 294 G1165 1170

30. HardwidgePR

Rodriguez-EscuderoI

GoodeD

DonohoeS

EngJ

2004 Proteomic analysis of the intestinal epithelial cell response to enteropathogenic Escherichia coli. J Biol Chem 279 20127 20136

31. MayerC

GrummtI

2006 Ribosome biogenesis and cell growth: mTOR coordinates transcription by all three classes of nuclear RNA polymerases. Oncogene 25 6384 6391

32. TsurugiK

MoritaT

OgataK

1974 Mode of degradation of ribosomes in regenerating rat liver in vivo. Eur J Biochem 45 119 126

33. LindstromMS

2009 Emerging functions of ribosomal proteins in gene-specific transcription and translation. Biochem Biophys Res Commun 379 167 170

34. MundyR

PetrovskaL

SmollettK

SimpsonN

WilsonRK

2004 Identification of a novel Citrobacter rodentium type III secreted protein, EspI, and roles of this and other secreted proteins in infection. Infect Immun 72 2288 2302

35. UgrinovaI

MonierK

IvaldiC

ThiryM

StorckS

2007 Inactivation of nucleolin leads to nucleolar disruption, cell cycle arrest and defects in centrosome duplication. BMC Mol Biol 8 66

36. MarescaM

MillerD

QuitardS

DeanP

KennyB

2005 Enteropathogenic Escherichia coli (EPEC) effector-mediated suppression of antimicrobial nitric oxide production in a small intestinal epithelial model system. Cell Microbiol 7 1749 1762

37. VaradiA

AinscowEK

AllanVJ

RutterGA

2002 Involvement of conventional kinesin in glucose-stimulated secretory granule movements and exocytosis in clonal pancreatic beta-cells. J Cell Sci 115 4177 4189

38. ChantretI

RodolosseA

BarbatA

DussaulxE

Brot-LarocheE

ZweibaumA

RoussetM

1994 Differential expression of sucrase-isomaltase in clones isolated from early and late passages of the cell line Caco-2: evidence for glucose-dependent negative regulation. J Cell Sci 107 (Pt 1) 213 225

39. LevineMM

BergquistEJ

NalinDR

WatermanDH

HornickRB

YoungCR

SotmanS

1978 Escherichia coli strains that cause diarrhoea but do not produce heat-labile or heat-stable enterotoxins and are non-invasive. Lancet 1 1119 1122

40. DonnenbergMS

KaperJB

1991 Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect Immun 59 4310 4317

41. WarawaJ

FinlayBB

KennyB

1999 Type III secretion-dependent hemolytic activity of enteropathogenic Escherichia coli. Infect Immun 67 5538 5540

42. KennyB

LaiLC

FinlayBB

DonnenbergMS

1996 EspA, a protein secreted by enteropathogenic Escherichia coli, is required to induce signals in epithelial cells. Mol Microbiol 20 313 323

43. KennyB

2001 The enterohaemorrhagic Escherichia coli (serotype O157:H7) Tir molecule is not functionally interchangeable for its enteropathogenic E. coli (serotype O127:H6) homologue. Cell Microbiol 3 499 510

44. AltoNM

WeflenAW

RardinMJ

YararD

LazarCS

TonikianR

KollerA

TaylorSS

BooneC

SidhuSS

SchmidSL

HechtGA

DixonJE

2007 The type III effector EspF coordinates membrane trafficking by the spatiotemporal activation of two eukaryotic signaling pathways. J Cell Biol 178 1265 1278

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

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PLOS Pathogens


2010 Číslo 6
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