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A Capsid-Encoded PPxY-Motif Facilitates Adenovirus Entry


Viruses use cellular machinery to enter and infect cells. In this study we address the cell entry mechanisms of nonenveloped adenoviruses (Ads). We show that protein VI, an internal capsid protein, is rapidly exposed after cell surface attachment and internalization and remains partially associated with the capsid during intracellular transport. We found that a PPxY motif within protein VI recruits Nedd4 E3 ubiquitin ligases to bind and ubiquitylate protein VI. We further show that this PPxY motif is involved in rapid, microtubule-dependent intracellular movement of protein VI. Ads with a mutated PPxY motif can efficiently escape endosomes but are defective in microtubule-dependent trafficking toward the nucleus. Likewise, depletion of Nedd4 ligases attenuates nuclear accumulation of incoming Ad particles and infection. Our data provide the first evidence that virus-encoded PPxY motifs are required during virus entry, which may be of significance for several other pathogens.


Vyšlo v časopise: A Capsid-Encoded PPxY-Motif Facilitates Adenovirus Entry. PLoS Pathog 6(3): e32767. doi:10.1371/journal.ppat.1000808
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1000808

Souhrn

Viruses use cellular machinery to enter and infect cells. In this study we address the cell entry mechanisms of nonenveloped adenoviruses (Ads). We show that protein VI, an internal capsid protein, is rapidly exposed after cell surface attachment and internalization and remains partially associated with the capsid during intracellular transport. We found that a PPxY motif within protein VI recruits Nedd4 E3 ubiquitin ligases to bind and ubiquitylate protein VI. We further show that this PPxY motif is involved in rapid, microtubule-dependent intracellular movement of protein VI. Ads with a mutated PPxY motif can efficiently escape endosomes but are defective in microtubule-dependent trafficking toward the nucleus. Likewise, depletion of Nedd4 ligases attenuates nuclear accumulation of incoming Ad particles and infection. Our data provide the first evidence that virus-encoded PPxY motifs are required during virus entry, which may be of significance for several other pathogens.


Zdroje

1. BrandenburgB

ZhuangX

2007 Virus trafficking - learning from single-virus tracking. Nat Rev Microbiol 5 197 208

2. GreberUF

WayM

2006 A superhighway to virus infection. Cell 124 741 754

3. LeopoldPL

CrystalRG

2007 Intracellular trafficking of adenovirus: many means to many ends. Adv Drug Deliv Rev 59 810 821

4. GreberUF

2002 Signalling in viral entry. Cell Mol Life Sci 59 608 626

5. LeopoldPL

PfisterKK

2006 Viral strategies for intracellular trafficking: motors and microtubules. Traffic 7 516 523

6. BaileyCJ

CrystalRG

LeopoldPL

2003 Association of adenovirus with the microtubule organizing center. J Virol 77 13275 13287

7. SuomalainenM

NakanoMY

KellerS

BouckeK

StidwillRP

1999 Microtubule-dependent plus- and minus end-directed motilities are competing processes for nuclear targeting of adenovirus. J Cell Biol 144 657 672

8. StrunzeS

TrotmanLC

BouckeK

GreberUF

2005 Nuclear targeting of adenovirus type 2 requires CRM1-mediated nuclear export. Mol Biol Cell 16 2999 3009

9. YuGY

LaiMM

2005 The ubiquitin-proteasome system facilitates the transfer of murine coronavirus from endosome to cytoplasm during virus entry. J Virol 79 644 648

10. KhorR

McElroyLJ

WhittakerGR

2003 The ubiquitin-vacuolar protein sorting system is selectively required during entry of influenza virus into host cells. Traffic 4 857 868

11. DuanD

YueY

YanZ

YangJ

EngelhardtJF

2000 Endosomal processing limits gene transfer to polarized airway epithelia by adeno-associated virus. J Clin Invest 105 1573 1587

12. YanZ

ZakR

LuxtonGW

RitchieTC

Bantel-SchaalU

2002 Ubiquitination of both adeno-associated virus type 2 and 5 capsid proteins affects the transduction efficiency of recombinant vectors. J Virol 76 2043 2053

13. BieniaszPD

2006 Late budding domains and host proteins in enveloped virus release. Virology 344 55 63

14. WilliamsRL

UrbeS

2007 The emerging shape of the ESCRT machinery. Nat Rev Mol Cell Biol 8 355 368

15. SaksenaS

WahlmanJ

TeisD

JohnsonAE

EmrSD

2009 Functional reconstitution of ESCRT-III assembly and disassembly. Cell 136 97 109

16. InghamRJ

GishG

PawsonT

2004 The Nedd4 family of E3 ubiquitin ligases: functional diversity within a common modular architecture. Oncogene 23 1972 1984

17. Shearwin-WhyattL

DaltonHE

FootN

KumarS

2006 Regulation of functional diversity within the Nedd4 family by accessory and adaptor proteins. Bioessays 28 617 628

18. UsamiY

PopovS

PopovaE

GottlingerHG

2008 Efficient and specific rescue of human immunodeficiency virus type 1 budding defects by a Nedd4-like ubiquitin ligase. J Virol 82 4898 4907

19. ChungHY

MoritaE

von SchwedlerU

MullerB

KrausslichHG

2008 NEDD4L overexpression rescues the release and infectivity of human immunodeficiency virus type 1 constructs lacking PTAP and YPXL late domains. J Virol 82 4884 4897

20. WirblichC

TanGS

PapaneriA

GodlewskiPJ

OrensteinJM

2008 PPEY motif within the rabies virus (RV) matrix protein is essential for efficient virion release and RV pathogenicity. J Virol 82 9730 9738

21. GalinierR

GoutE

Lortat-JacobH

WoodJ

ChroboczekJ

2002 Adenovirus protein involved in virus internalization recruits ubiquitin-protein ligases. Biochemistry 41 14299 14305

22. ZhangY

BergelsonJM

2005 Adenovirus receptors. J Virol 79 12125 12131

23. MeierO

GreberUF

2004 Adenovirus endocytosis. J Gene Med 6 Suppl 1 S152 163

24. RaumaT

TuukkanenJ

BergelsonJM

DenningG

HautalaT

1999 rab5 GTPase regulates adenovirus endocytosis. J Virol 73 9664 9668

25. WickhamTJ

MathiasP

ChereshDA

NemerowGR

1993 Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. Cell 73 309 319

26. GreberUF

WillettsM

WebsterP

HeleniusA

1993 Stepwise dismantling of adenovirus 2 during entry into cells. Cell 75 477 486

27. GreberUF

WebsterP

WeberJ

HeleniusA

1996 The role of the adenovirus protease on virus entry into cells. Embo J 15 1766 1777

28. WiethoffCM

WodrichH

GeraceL

NemerowGR

2005 Adenovirus protein VI mediates membrane disruption following capsid disassembly. J Virol 79 1992 2000

29. SaphireAC

GuanT

SchirmerEC

NemerowGR

GeraceL

2000 Nuclear import of adenovirus DNA in vitro involves the nuclear protein import pathway and hsc70. J Biol Chem 275 4298 4304

30. TrotmanLC

MosbergerN

FornerodM

StidwillRP

GreberUF

2001 Import of adenovirus DNA involves the nuclear pore complex receptor CAN/Nup214 and histone H1. Nat Cell Biol 3 1092 1100

31. WodrichH

CassanyA

D'AngeloMA

GuanT

NemerowG

2006 Adenovirus core protein pVII is translocated into the nucleus by multiple import receptor pathways. J Virol 80 9608 9618

32. LoktevAV

ZhangQ

BeckJS

SearbyCC

ScheetzTE

2008 A BBSome subunit links ciliogenesis, microtubule stability, and acetylation. Dev Cell 15 854 865

33. CopelandNG

JenkinsNA

CourtDL

2001 Recombineering: a powerful new tool for mouse functional genomics. Nat Rev Genet 2 769 779

34. RuzsicsZ

WagnerM

OsterlehnerA

CookJ

KoszinowskiU

2006 Transposon-assisted cloning and traceless mutagenesis of adenoviruses: Development of a novel vector based on species D. J Virol 80 8100 8113

35. YasudaJ

HunterE

1998 A proline-rich motif (PPPY) in the Gag polyprotein of Mason-Pfizer monkey virus plays a maturation-independent role in virion release. J Virol 72 4095 4103

36. GottweinE

BodemJ

MullerB

SchmechelA

ZentgrafH

2003 The Mason-Pfizer monkey virus PPPY and PSAP motifs both contribute to virus release. J Virol 77 9474 9485

37. WodrichH

GuanT

CingolaniG

Von SeggernD

NemerowG

2003 Switch from capsid protein import to adenovirus assembly by cleavage of nuclear transport signals. Embo J 22 6245 6255

38. Martin-SerranoJ

EastmanSW

ChungW

BieniaszPD

2005 HECT ubiquitin ligases link viral and cellular PPXY motifs to the vacuolar protein-sorting pathway. J Cell Biol 168 89 101

39. GastaldelliM

ImelliN

BouckeK

AmstutzB

MeierO

2008 Infectious Adenovirus Type 2 Transport Through Early but not Late Endosomes. Traffic

40. Medina-KauweLK

2003 Endocytosis of adenovirus and adenovirus capsid proteins. Adv Drug Deliv Rev 55 1485 1496

41. SvenssonU

1985 Role of vesicles during adenovirus 2 internalization into HeLa cells. J Virol 55 442 449

42. NakanoMY

BouckeK

SuomalainenM

StidwillRP

GreberUF

2000 The first step of adenovirus type 2 disassembly occurs at the cell surface, independently of endocytosis and escape to the cytosol. J Virol 74 7085 7095

43. SabanSD

SilvestryM

NemerowGR

StewartPL

2006 Visualization of alpha-helices in a 6-angstrom resolution cryoelectron microscopy structure of adenovirus allows refinement of capsid protein assignments. J Virol 80 12049 12059

44. RussellWC

2009 Adenoviruses: update on structure and function. J Gen Virol 90 1 20

45. FabryCM

Rosa-CalatravaM

ConwayJF

ZubietaC

CusackS

2005 A quasi-atomic model of human adenovirus type 5 capsid. Embo J 24 1645 1654

46. NeumannG

EbiharaH

TakadaA

NodaT

KobasaD

2005 Ebola virus VP40 late domains are not essential for viral replication in cell culture. J Virol 79 10300 10307

47. MoritaE

SandrinV

ChungHY

MorhamSG

GygiSP

2007 Human ESCRT and ALIX proteins interact with proteins of the midbody and function in cytokinesis. Embo J 26 4215 4227

48. FejerG

DrechselL

LieseJ

SchleicherU

RuzsicsZ

2008 Key role of splenic myeloid DCs in the IFN-alphabeta response to adenoviruses in vivo. PLoS Pathog 4 e1000208 doi:10.1371/journal.ppat.1000208

49. ChroboczekJ

GoutE

FavierAL

GalinierR

2003 Novel partner proteins of adenovirus penton. Curr Top Microbiol Immunol 272 37 55

50. SalinasS

BilslandLG

HenaffD

WestonAE

KerielA

2009 CAR-associated vesicular transport of an adenovirus in motor neuron axons. PLoS Pathog 5 e1000442

51. MitterederN

MarchKL

TrapnellBC

1996 Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy. J Virol 70 7498 7509

52. Segura-MoralesC

PesciaC

Chatellard-CausseC

SadoulR

BertrandE

2005 Tsg101 and Alix interact with murine leukemia virus Gag and cooperate with Nedd4 ubiquitin ligases during budding. J Biol Chem 280 27004 27012

53. FotiaAB

EkbergJ

AdamsDJ

CookDI

PoronnikP

2004 Regulation of neuronal voltage-gated sodium channels by the ubiquitin-protein ligases Nedd4 and Nedd4-2. J Biol Chem 279 28930 28935

54. RossiM

AqeilanRI

NealeM

CandiE

SalomoniP

2006 The E3 ubiquitin ligase Itch controls the protein stability of p63. Proc Natl Acad Sci U S A 103 12753 12758

55. BubeckA

WagnerM

RuzsicsZ

LotzerichM

IglesiasM

2004 Comprehensive mutational analysis of a herpesvirus gene in the viral genome context reveals a region essential for virus replication. J Virol 78(15) 8026 8035

56. CherepanovPP

WackernagelW

1995 Gene disruption in Escherichia coli: TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. Gene 158(1) 9 14

57. WarmingS

CostantinoN

CourtDL

JenkinsNA

CopelandNG

2005 Simple and highly efficient BAC recombineering using galK selection. Nucleic Acids Res 33(4) e36

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Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

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PLOS Pathogens


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