#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Trafficking of Hepatitis C Virus Core Protein during Virus Particle Assembly


Hepatitis C virus (HCV) core protein is directed to the surface of lipid droplets (LD), a step that is essential for infectious virus production. However, the process by which core is recruited from LD into nascent virus particles is not well understood. To investigate the kinetics of core trafficking, we developed methods to image functional core protein in live, virus-producing cells. During the peak of virus assembly, core formed polarized caps on large, immotile LDs, adjacent to putative sites of assembly. In addition, LD-independent, motile puncta of core were found to traffic along microtubules. Importantly, core was recruited from LDs into these puncta, and interaction between the viral NS2 and NS3-4A proteins was essential for this recruitment process. These data reveal new aspects of core trafficking and identify a novel role for viral nonstructural proteins in virus particle assembly.


Vyšlo v časopise: Trafficking of Hepatitis C Virus Core Protein during Virus Particle Assembly. PLoS Pathog 7(10): e32767. doi:10.1371/journal.ppat.1002302
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002302

Souhrn

Hepatitis C virus (HCV) core protein is directed to the surface of lipid droplets (LD), a step that is essential for infectious virus production. However, the process by which core is recruited from LD into nascent virus particles is not well understood. To investigate the kinetics of core trafficking, we developed methods to image functional core protein in live, virus-producing cells. During the peak of virus assembly, core formed polarized caps on large, immotile LDs, adjacent to putative sites of assembly. In addition, LD-independent, motile puncta of core were found to traffic along microtubules. Importantly, core was recruited from LDs into these puncta, and interaction between the viral NS2 and NS3-4A proteins was essential for this recruitment process. These data reveal new aspects of core trafficking and identify a novel role for viral nonstructural proteins in virus particle assembly.


Zdroje

1. LindenbachBDThielHJRiceCM 2007 Flaviviridae: The viruses and their replication. KnipeDMHowleyPM Fields Virology Fifth ed. Philadelphia Lippincott-Raven Publishers 1101 1152

2. JonesDMPatelAHTargett-AdamsPMcLauchlanJ 2009 The hepatitis C virus NS4B protein can trans-complement viral RNA replication and modulates production of infectious virus. J Virol 83 2163 2177

3. MurrayCLJonesCTRiceCM 2008 Architects of assembly: roles of Flaviviridae non-structural proteins in virion morphogenesis. Nat Rev Microbiol 6 699 708

4. PhanTKohlwayADimberuPPyleAMLindenbachBD 2011 The acidic domain of hepatitis C virus NS4A contributes to RNA replication and virus particle assembly. J Virol 85 1193 1204

5. McLauchlanJ 2000 Properties of the hepatitis C virus core protein: a structural protein that modulates cellular processes. J Viral Hepat 7 2 14

6. BoulantSVanbelleCEbelCPeninFLavergneJP 2005 Hepatitis C virus core protein is a dimeric alpha-helical protein exhibiting membrane protein features. J Virol 79 11353 11365

7. BoulantSMontserretRHopeRGRatinierMTargett-AdamsP 2006 Structural determinants that target the hepatitis C virus core protein to lipid droplets. J Biol Chem 281 22236 22247

8. KoppMMurrayCLJonesCTRiceCM 2010 Genetic analysis of the carboxy-terminal region of the hepatitis C virus core protein. J Virol 84 1666 1673

9. OkamotoKMoriYKomodaYOkamotoTOkochiM 2008 Intramembrane processing by signal peptide peptidase regulates the membrane localization of hepatitis C virus core protein and viral propagation. J Virol 82 8349 8361

10. HüssyPLangenHMousJJacobsenH 1996 Hepatitis C virus core protein: carboxy-terminal boundaries of two processed species suggest cleavage by a signal peptide peptidase. Virology 224 93 104

11. BarbaGHarperFHaradaTKoharaMGoulinetS 1997 Hepatitis C virus core protein shows a cytoplasmic localization and associates to cellular lipid storage droplets. Proc Natl Acad Sci USA 94 1200 1205

12. McLauchlanJLembergMKHopeGMartoglioB 2002 Intramembrane proteolysis promotes trafficking of hepatitis C virus core protein to lipid droplets. EMBO J 21 3980 3988

13. MoradpourDEnglertCWakitaTWandsJR 1996 Characterization of cell lines allowing tightly regulated expression of hepatitis C virus core protein. Virology 222 51 63

14. BellerMThielKThulPJJackleH 2010 Lipid droplets: a dynamic organelle moves into focus. FEBS Lett 584 2176 2182

15. FareseRVJrWaltherTC 2009 Lipid droplets finally get a little R-E-S-P-E-C-T. Cell 139 855 860

16. HerkerEHarrisCHernandezCCarpentierAKaehlckeK 2010 Efficient hepatitis C virus particle formation requires diacylglycerol acyltransferase-1. Nat Med 16 1295 1298

17. BrasaemleDL 2007 Thematic review series: adipocyte biology. The perilipin family of structural lipid droplet proteins: stabilization of lipid droplets and control of lipolysis. J Lipid Res 48 2547 2559

18. FujimotoYItabeHSakaiJMakitaMNodaJ 2004 Identification of major proteins in the lipid droplet-enriched fraction isolated from the human hepatocyte cell line HuH7. Biochim Biophys Acta 1644 47 59

19. MartinSPartonRG 2008 Characterization of Rab18, a lipid droplet-associated small GTPase. Methods Enzymol 438 109 129

20. OzekiSChengJTauchi-SatoKHatanoNTaniguchiH 2005 Rab18 localizes to lipid droplets and induces their close apposition to the endoplasmic reticulum-derived membrane. J Cell Sci 118 2601 2611

21. TurroSIngelmo-TorresMEstanyolJMTebarFFernandezMA 2006 Identification and characterization of associated with lipid droplet protein 1: A novel membrane-associated protein that resides on hepatic lipid droplets. Traffic 7 1254 1269

22. MiyanariYAtsuzawaKUsudaNWatashiKHishikiT 2007 The lipid droplet is an important organelle for hepatitis C virus production. Nat Cell Biol 9 1089 1097

23. AppelNZayasMMillerSKrijnse-LockerJSchallerT 2008 Essential role of domain III of nonstructural protein 5A for hepatitis C virus infectious particle assembly. PLoS Pathog 4 e1000035

24. BoulantSTargett-AdamsPMcLauchlanJ 2007 Disrupting the association of hepatitis C virus core protein with lipid droplets correlates with a loss in production of infectious virus. J Gen Virol 88 2204 2213

25. MasakiTSuzukiRMurakamiKAizakiHIshiiK 2008 Interaction of hepatitis C virus nonstructural protein 5A with core protein is critical for the production of infectious virus particles. J Virol 82 7964 7976

26. ShavinskayaABoulantSPeninFMcLauchlanJBartenschlagerR 2007 The lipid droplet binding domain of hepatitis C virus core protein is a major determinant for efficient virus assembly. J Biol Chem 282 37158 37169

27. CocquerelLDuvetSMeunierJCPillezACacanR 1999 The transmembrane domain of hepatitis C virus glycoprotein E1 is a signal for static retention in the endoplasmic reticulum. J Virol 73 2641 2649

28. CocquerelLMeunierJCPillezAWychowskiCDubuissonJ 1998 A retention signal necessary and sufficient for endoplasmic reticulum localization maps to the transmembrane domain of hepatitis C virus glycoprotein E2. J Virol 72 2183 2191

29. HelleFVieyresGElkriefLPopescuCIWychowskiC 2010 Role of N-linked glycans in the functions of hepatitis C virus envelope proteins incorporated into infectious virions. J Virol 84 11905 11915

30. GastaminzaPKapadiaSBChisariFV 2006 Differential biophysical properties of infectious intracellular and secreted hepatitis C virus particles. J Virol 80 11074 11081

31. MukhopadhyaySKuhnRJRossmannMG 2005 A structural perspective of the flavivirus life cycle. Nat Rev Microbiol 3 13 22

32. MagnussonBAspLBostromPRuizMStillemark-BilltonP 2006 Adipocyte differentiation-related protein promotes fatty acid storage in cytosolic triglycerides and inhibits secretion of very low-density lipoproteins. Arterioscler Thromb Vasc Biol 26 1566 1571

33. MartinSDriessenKNixonSJZerialMPartonRG 2005 Regulated localization of Rab18 to lipid droplets: effects of lipolytic stimulation and inhibition of lipid droplet catabolism. J Biol Chem 280 42325 42335

34. BartenschlagerRPeninFLohmannVAndreP 2011 Assembly of infectious hepatitis C virus particles. Trends Microbiol 19 95 103

35. NoppornpanthSSmitsSLLienTXPoovorawanYOsterhausAD 2007 Characterization of hepatitis C virus deletion mutants circulating in chronically infected patients. J Virol 81 12496 12503

36. KlausnerRDDonaldsonJGLippincott-SchwartzJ 1992 Brefeldin A: insights into the control of membrane traffic and organelle structure. J Cell Biol 116 1071 1080

37. GoueslainLAlsalehKHorellouPRoingeardPDescampsV 2010 Identification of GBF1 as a cellular factor required for hepatitis C virus RNA replication. J Virol 84 773 787

38. GastaminzaPChengGWielandSZhongJLiaoW 2008 Cellular determinants of hepatitis C virus assembly, maturation, degradation, and secretion. J Virol 82 2120 2129

39. BellerMSztalrydCSouthallNBellMJackleH 2008 COPI complex is a regulator of lipid homeostasis. PLoS Biol 6 e292

40. GuoYWaltherTCRaoMStuurmanNGoshimaG 2008 Functional genomic screen reveals genes involved in lipid-droplet formation and utilization. Nature 453 657 661

41. SoniKGMardonesGASougratRSmirnovaEJacksonCL 2009 Coatomer-dependent protein delivery to lipid droplets. J Cell Sci 122 1834 1841

42. GaiettaGDeerinckTJAdamsSRBouwerJTourO 2002 Multicolor and electron microscopic imaging of connexin trafficking. Science 296 503 507

43. JiraskoVMontserretRLeeJYGouttenoireJMoradpourD 2010 Structural and functional studies of nonstructural protein 2 of the hepatitis C virus reveal its key role as organizer of virion assembly. PLoS Path 6 e1001233

44. MaYAnantpadmaMTimpeJMShanmugamSSinghSM 2011 Hepatitis C virus NS2 protein serves as a scaffold for virus assembly by interacting with both structural and nonstructural proteins. J Virol 85 86 97

45. StaplefordKALindenbachBD 2011 Hepatitis C Virus NS2 Coordinates Virus Particle Assembly through Physical Interactions with the E1-E2 Glycoprotein and NS3-NS4A Enzyme Complexes. J Virol 85 1706 1717

46. PhanTBeranRKPetersCLorenzICLindenbachBD 2009 Hepatitis C virus NS2 protein contributes to virus particle assembly via opposing epistatic interactions with the E1-E2 glycoprotein and NS3-NS4A enzyme complexes. J Virol 83 8379 8395

47. OhsakiYChengJFujitaATokumotoTFujimotoT 2006 Cytoplasmic lipid droplets are sites of convergence of proteasomal and autophagic degradation of apolipoprotein B. Mol Biol Cell 17 2674 2683

48. GriffinBAAdamsSRJonesJTsienRY 2000 Fluorescent labeling of recombinant proteins in living cells with FlAsH. Methods Enzymol 327 565 578

49. BoulantSDouglasMWMoodyLBudkowskaATargett-AdamsP 2008 Hepatitis C virus core protein induces lipid droplet redistribution in a microtubule- and Dynein-dependent manner. Traffic 9 1268 1282

50. LynRKKennedyDCStolowARidsdaleAPezackiJP 2010 Dynamics of lipid droplets induced by the hepatitis C virus core protein. Bioch Biophys Res Comm 399 518 524

51. LaiCKJengKSMachidaKLaiMM 2010 Hepatitis C virus egress and release depend on endosomal trafficking of core protein. J Virol 84 11590 11598

52. Targett-AdamsPChambersDGledhillSHopeRGCoyJF 2003 Live cell analysis and targeting of the lipid droplet-binding adipocyte differentiation-related protein. J Biol Chem 278 15998 16007

53. ShanerNCLinMZMcKeownMRSteinbachPAHazelwoodKL 2008 Improving the photostability of bright monomeric orange and red fluorescent proteins. Nat Methods 5 545 551

54. BlightKJMcKeatingJARiceCM 2002 Highly permissive cell lines for subgenomic and genomic hepatitis C virus RNA replication. J Virol 76 13001 13014

55. LawMMaruyamaTLewisJGiangETarrAW 2008 Broadly neutralizing antibodies protect against hepatitis C virus quasispecies challenge. Nat Med 14 25 27

56. LindenbachBDEvansMJSyderAJWolkBTellinghuisenTL 2005 Complete replication of hepatitis C virus in cell culture. Science 309 623 626

57. LindenbachBD 2009 Measuring HCV infectivity produced in cell culture and in vivo. Methods Mol Biol 510 329 336

58. FrenchAPMillsSSwarupRBennettMJPridmoreTP 2008 Colocalization of fluorescent markers in confocal microscope images of plant cells. Nat Protoc 3 619 628

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2011 Číslo 10
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

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#