#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

The Length of Vesicular Stomatitis Virus Particles Dictates a Need for Actin Assembly during Clathrin-Dependent Endocytosis


Microbial pathogens exploit the clathrin endocytic machinery to enter host cells. Vesicular stomatitis virus (VSV), an enveloped virus with bullet-shaped virions that measure 70×200 nm, enters cells by clathrin-dependent endocytosis. We showed previously that VSV particles exceed the capacity of typical clathrin-coated vesicles and instead enter through endocytic carriers that acquire a partial clathrin coat and require local actin filament assembly to complete vesicle budding and internalization. To understand why the actin system is required for VSV uptake, we compared the internalization mechanisms of VSV and its shorter (75 nm long) defective interfering particle, DI-T. By imaging the uptake of individual particles into live cells, we found that, as with parental virions, DI-T enters via the clathrin endocytic pathway. Unlike VSV, DI-T internalization occurs through complete clathrin-coated vesicles and does not require actin polymerization. Since VSV and DI-T particles display similar surface densities of the same attachment glycoprotein, we conclude that the physical properties of the particle dictate whether a virus-containing clathrin pit engages the actin system. We suggest that the elongated shape of a VSV particle prevents full enclosure by the clathrin coat and that stalling of coat assembly triggers recruitment of the actin machinery to finish the internalization process. Since some enveloped viruses have pleomorphic particle shapes and sizes, our work suggests that they may use altered modes of endocytic uptake. More generally, our findings show the importance of cargo geometry for specifying cellular entry modes, even when the receptor recognition properties of a ligand are maintained.


Vyšlo v časopise: The Length of Vesicular Stomatitis Virus Particles Dictates a Need for Actin Assembly during Clathrin-Dependent Endocytosis. PLoS Pathog 6(9): e32767. doi:10.1371/journal.ppat.1001127
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1001127

Souhrn

Microbial pathogens exploit the clathrin endocytic machinery to enter host cells. Vesicular stomatitis virus (VSV), an enveloped virus with bullet-shaped virions that measure 70×200 nm, enters cells by clathrin-dependent endocytosis. We showed previously that VSV particles exceed the capacity of typical clathrin-coated vesicles and instead enter through endocytic carriers that acquire a partial clathrin coat and require local actin filament assembly to complete vesicle budding and internalization. To understand why the actin system is required for VSV uptake, we compared the internalization mechanisms of VSV and its shorter (75 nm long) defective interfering particle, DI-T. By imaging the uptake of individual particles into live cells, we found that, as with parental virions, DI-T enters via the clathrin endocytic pathway. Unlike VSV, DI-T internalization occurs through complete clathrin-coated vesicles and does not require actin polymerization. Since VSV and DI-T particles display similar surface densities of the same attachment glycoprotein, we conclude that the physical properties of the particle dictate whether a virus-containing clathrin pit engages the actin system. We suggest that the elongated shape of a VSV particle prevents full enclosure by the clathrin coat and that stalling of coat assembly triggers recruitment of the actin machinery to finish the internalization process. Since some enveloped viruses have pleomorphic particle shapes and sizes, our work suggests that they may use altered modes of endocytic uptake. More generally, our findings show the importance of cargo geometry for specifying cellular entry modes, even when the receptor recognition properties of a ligand are maintained.


Zdroje

1. ConnerSD

SchmidSL

2003 Regulated portals of entry into the cell. Nature 422 37 44

2. KirchhausenT

2000 Clathrin. Annu Rev Biochem 69 699 727

3. KirchhausenT

2009 Imaging endocytic clathrin structures in living cells. Trends Cell Biol 19 596 605

4. OwenDJ

EvansPR

1998 A structural explanation for the recognition of tyrosine-based endocytotic signals. Science 282 1327 1332

5. HoningS

RicottaD

KraussM

SpateK

SpolaoreB

2005 Phosphatidylinositol-(4,5)-bisphosphate regulates sorting signal recognition by the clathrin-associated adaptor complex AP2. Mol Cell 18 519 531

6. DamkeH

BabaT

WarnockDE

SchmidSL

1994 Induction of mutant dynamin specifically blocks endocytic coated vesicle formation. J Cell Biol 127 915 934

7. LeeDW

ZhaoX

ZhangF

EisenbergE

GreeneLE

2005 Depletion of GAK/auxilin 2 inhibits receptor-mediated endocytosis and recruitment of both clathrin and clathrin adaptors. J Cell Sci 118 4311 4321

8. MassolRH

BollW

GriffinAM

KirchhausenT

2006 A burst of auxilin recruitment determines the onset of clathrin-coated vesicle uncoating. Proc Natl Acad Sci U S A 103 10265 10270

9. EhrlichM

BollW

Van OijenA

HariharanR

ChandranK

2004 Endocytosis by random initiation and stabilization of clathrin-coated pits. Cell 118 591 605

10. LoerkeD

MettlenM

YararD

JaqamanK

JaqamanH

2009 Cargo and dynamin regulate clathrin-coated pit maturation. PLoS Biol 7 e57

11. HanoverJA

BeguinotL

WillinghamMC

PastanIH

1985 Transit of receptors for epidermal growth factor and transferrin through clathrin-coated pits. Analysis of the kinetics of receptor entry. J Biol Chem 260 15938 15945

12. AndersonRG

BrownMS

GoldsteinJL

1977 Role of the coated endocytic vesicle in the uptake of receptor-bound low density lipoprotein in human fibroblasts. Cell 10 351 364

13. van der SchaarHM

RustMJ

ChenC

van der Ende-MetselaarH

WilschutJ

2008 Dissecting the cell entry pathway of dengue virus by single-particle tracking in living cells. PLoS Pathog 4 e1000244

14. RustMJ

LakadamyaliM

ZhangF

ZhuangX

2004 Assembly of endocytic machinery around individual influenza viruses during viral entry. Nat Struct Mol Biol 11 567 573

15. CuretonDK

MassolRH

SaffarianS

KirchhausenTL

WhelanSP

2009 Vesicular stomatitis virus enters cells through vesicles incompletely coated with clathrin that depend upon actin for internalization. PLoS Pathog 5 e1000394

16. VeigaE

GuttmanJA

BonazziM

BoucrotE

Toledo-AranaA

2007 Invasive and adherent bacterial pathogens co-Opt host clathrin for infection. Cell Host Microbe 2 340 351

17. SimpsonRW

HauserRE

DalesS

1969 Viropexis of vesicular stomatitis virus by L cells. Virology 37 285 290

18. HarrisA

CardoneG

WinklerDC

HeymannJB

BrecherM

2006 Influenza virus pleiomorphy characterized by cryoelectron tomography. Proc Natl Acad Sci U S A 103 19123 19127

19. MeierE

HarmisonGG

KeeneJD

SchubertM

1984 Sites of copy choice replication involved in generation of vesicular stomatitis virus defective-interfering particle RNAs. J Virol 51 515 521

20. HuangAS

WagnerRR

1966 Defective T particles of vesicular stomatitis virus. II. Biologic role in homologous interference. Virology 30 173 181

21. PengG

TsaoJ

ScheinS

GreenTJ

LuoM

2010 Cryo-EM model of the bullet-shaped vesicular stomatitis virus. Science 327 689 693

22. HuangAS

GreenawaltJW

WagnerRR

1966 Defective T particles of vesicular stomatitis virus. I. Preparation, morphology, and some biologic properties. Virology 30 161 172

23. WagnerRR

SchnaitmanTA

SnyderRM

1969 Structural proteins of vesicular stomatitis viruses. J Virol 3 395 403

24. PattnaikAK

BallLA

LeGroneAW

WertzGW

1992 Infectious defective interfering particles of VSV from transcripts of a cDNA clone. Cell 69 1011 1020

25. WhelanSP

BallLA

BarrJN

WertzGT

1995 Efficient recovery of infectious vesicular stomatitis virus entirely from cDNA clones. Proc Natl Acad Sci U S A 92 8388 8392

26. SaffarianS

CocucciE

KirchhausenT

2009 Distinct dynamics of endocytic clathrin-coated pits and coated plaques. PLoS Biol 7 e1000191

27. UrunoT

LiuJ

ZhangP

FanY

EgileC

2001 Activation of Arp2/3 complex-mediated actin polymerization by cortactin. Nat Cell Biol 3 259 266

28. McNivenMA

KimL

KruegerEW

OrthJD

CaoH

2000 Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape. J Cell Biol 151 187 198

29. MerrifieldCJ

PerraisD

ZenisekD

2005 Coupling between clathrin-coated-pit invagination, cortactin recruitment, and membrane scission observed in live cells. Cell 121 593 606

30. CoueM

BrennerSL

SpectorI

KornED

1987 Inhibition of actin polymerization by latrunculin A. FEBS Lett 213 316 318

31. CoyneCB

BergelsonJM

2006 Virus-induced Abl and Fyn kinase signals permit coxsackievirus entry through epithelial tight junctions. Cell 124 119 131

32. CoyneCB

KimKS

BergelsonJM

2007 Poliovirus entry into human brain microvascular cells requires receptor-induced activation of SHP-2. Embo J 26 4016 4028

33. KaksonenM

ToretCP

DrubinDG

2006 Harnessing actin dynamics for clathrin-mediated endocytosis. Nat Rev Mol Cell Biol 7 404 414

34. KaksonenM

SunY

DrubinDG

2003 A pathway for association of receptors, adaptors, and actin during endocytic internalization. Cell 115 475 487

35. IdrissiFZ

GrotschH

Fernandez-GolbanoIM

Presciatto-BaschongC

RiezmanH

2008 Distinct acto/myosin-I structures associate with endocytic profiles at the plasma membrane. J Cell Biol 180 1219 1232

36. HeuserJ

1980 Three-dimensional visualization of coated vesicle formation in fibroblasts. J Cell Biol 84 560 583

37. GottliebTA

IvanovIE

AdesnikM

SabatiniDD

1993 Actin microfilaments play a critical role in endocytosis at the apical but not the basolateral surface of polarized epithelial cells. J Cell Biol 120 695 710

38. Da CostaSR

SouE

XieJ

YarberFA

OkamotoCT

2003 Impairing actin filament or syndapin functions promotes accumulation of clathrin-coated vesicles at the apical plasma membrane of acinar epithelial cells. Mol Biol Cell 14 4397 4413

39. ShupliakovO

BloomO

GustafssonJS

KjaerulffO

LowP

2002 Impaired recycling of synaptic vesicles after acute perturbation of the presynaptic actin cytoskeleton. Proc Natl Acad Sci U S A 99 14476 14481

40. BourneJ

MorganJR

PieriboneVA

2006 Actin polymerization regulates clathrin coat maturation during early stages of synaptic vesicle recycling at lamprey synapses. J Comp Neurol 497 600 609

41. BoucrotE

SaffarianS

MassolR

KirchhausenT

EhrlichM

2006 Role of lipids and actin in the formation of clathrin-coated pits. Exp Cell Res 312 4036 4048

42. QualmannB

RoosJ

DiGregorioPJ

KellyRB

1999 Syndapin I, a synaptic dynamin-binding protein that associates with the neural Wiskott-Aldrich syndrome protein. Mol Biol Cell 10 501 513

43. SouletF

YararD

LeonardM

SchmidSL

2005 SNX9 regulates dynamin assembly and is required for efficient clathrin-mediated endocytosis. Mol Biol Cell 16 2058 2067

44. YararD

Waterman-StorerCM

SchmidSL

2007 SNX9 couples actin assembly to phosphoinositide signals and is required for membrane remodeling during endocytosis. Dev Cell 13 43 56

45. RingstadN

NemotoY

De CamilliP

1997 The SH3p4/Sh3p8/SH3p13 protein family: binding partners for synaptojanin and dynamin via a Grb2-like Src homology 3 domain. Proc Natl Acad Sci U S A 94 8569 8574

46. OtsukiM

ItohT

TakenawaT

2003 Neural Wiskott-Aldrich syndrome protein is recruited to rafts and associates with endophilin A in response to epidermal growth factor. J Biol Chem 278 6461 6469

47. DawsonJC

LeggJA

MacheskyLM

2006 Bar domain proteins: a role in tubulation, scission and actin assembly in clathrin-mediated endocytosis. Trends Cell Biol 16 493 498

48. RobertsPC

CompansRW

1998 Host cell dependence of viral morphology. Proc Natl Acad Sci U S A 95 5746 5751

49. RobertsPC

LambRA

CompansRW

1998 The M1 and M2 proteins of influenza A virus are important determinants in filamentous particle formation. Virology 240 127 137

50. NeumanBW

AdairBD

BurnsJW

MilliganRA

BuchmeierMJ

2005 Complementarity in the supramolecular design of arenaviruses and retroviruses revealed by electron cryomicroscopy and image analysis. J Virol 79 3822 3830

51. MurphyFA

WebbPA

JohnsonKM

WhitfieldSG

ChappellWA

1970 Arenoviruses in Vero cells: ultrastructural studies. J Virol 6 507 518

52. RojekJM

SanchezAB

NguyenNT

de la TorreJC

KunzS

2008 Different mechanisms of cell entry by human-pathogenic Old World and New World arenaviruses. J Virol 82 7677 7687

53. ChandranK

SullivanNJ

FelborU

WhelanSP

CunninghamJM

2005 Endosomal proteolysis of the Ebola virus glycoprotein is necessary for infection. Science 308 1643 1645

54. NegreteOA

LevroneyEL

AguilarHC

Bertolotti-CiarletA

NazarianR

2005 EphrinB2 is the entry receptor for Nipah virus, an emergent deadly paramyxovirus. Nature 436 401 405

55. ClementeR

de la TorreJC

2009 Cell entry of Borna disease virus follows a clathrin-mediated endocytosis pathway that requires Rab5 and microtubules. J Virol 83 10406 10416

56. BhattacharyyaS

WarfieldKL

RuthelG

BavariS

AmanMJ

2010 Ebola virus uses clathrin-mediated endocytosis as an entry pathway. Virology 401 18 28

57. Le ClaincheC

PaulyBS

ZhangCX

Engqvist-GoldsteinAE

CunninghamK

2007 A Hip1R-cortactin complex negatively regulates actin assembly associated with endocytosis. Embo J 26 1199 1210

58. KaksonenM

PengHB

RauvalaH

2000 Association of cortactin with dynamic actin in lamellipodia and on endosomal vesicles. J Cell Sci 113 Pt 24 4421 4426

59. MaupinP

PollardTD

1983 Improved preservation and staining of HeLa cell actin filaments, clathrin-coated membranes, and other cytoplasmic structures by tannic acid-glutaraldehyde-saponin fixation. J Cell Biol 96 51 62

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2010 Číslo 9
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Aktuální možnosti diagnostiky a léčby litiáz
nový kurz
Autori: MUDr. Tomáš Ürge, PhD.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#