A Cysteine Protease Inhibitor of Is Essential for Exo-erythrocytic Development

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Coordinated protease activity is essential to parasite survival. Throughout its life cycle, the Plasmodium parasite expresses a potent cysteine protease inhibitor that has the potential to inhibit parasite as well as host cell cysteine proteases. We have generated a stage-specific knockout of this inhibitor and were able to analyze its function in all life cycle stages. Interestingly, although constitutively expressed, the inhibitor primarily appears to play an important role in sporozoite gliding, liver stage development and egress from hepatocytes whereas blood stage parasites lacking the inhibitor exhibited only mild attenuation. Parasite sexual stage development was not affected and development continued normally within the mosquito. However, sporozoites lacking the inhibitor show a strong phenotype; they are completely blocked in motility and thus cannot transmigrate or invade cells. Complementation of knockout parasites by exogenous expression of the inhibitor completely restored parasite virulence.

Vyšlo v časopise: A Cysteine Protease Inhibitor of Is Essential for Exo-erythrocytic Development. PLoS Pathog 10(8): e32767. doi:10.1371/journal.ppat.1004336
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004336

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1. BrightAT, WinzelerEA (2013) Epidemiology: resistance mapping in malaria. Nature 498: 446–447.

2. ChaijaroenkulW, BangchangKN, MungthinM, WardSA (2005) In vitro antimalarial drug susceptibility in Thai border areas from 1998–2003. Malaria journal 4: 37.

3. WilairatanaP, KrudsoodS, TreeprasertsukS, ChalermrutK, LooareesuwanS (2002) The future outlook of antimalarial drugs and recent work on the treatment of malaria. Archives of medical research 33: 416–421.

4. BinderEM, KimK (2004) Location, location, location: trafficking and function of secreted proteases of Toxoplasma and Plasmodium. Traffic 5: 914–924.

5. BlackmanMJ (2008) Malarial proteases and host cell egress: an ‘emerging’ cascade. Cell Microbiol 10: 1925–1934.

6. CoppensI, SullivanDJ, PriggeST (2010) An update on the rapid advances in malaria parasite cell biology. Trends in parasitology 26: 305–310.

7. DowseTJ, KoussisK, BlackmanMJ, Soldati-FavreD (2008) Roles of proteases during invasion and egress by Plasmodium and Toxoplasma. Sub-cellular biochemistry 47: 121–139.

8. DrewME, BanerjeeR, UffmanEW, GilbertsonS, RosenthalPJ, et al. (2008) Plasmodium food vacuole plasmepsins are activated by falcipains. The Journal of biological chemistry 283: 12870–12876.

9. GoldbergDE (2013) Complex nature of malaria parasite hemoglobin degradation [corrected]. Proceedings of the National Academy of Sciences of the United States of America 110: 5283–5284.

10. HoggT, NagarajanK, HerzbergS, ChenL, ShenX, et al. (2006) Structural and functional characterization of Falcipain-2, a hemoglobinase from the malarial parasite Plasmodium falciparum. The Journal of biological chemistry 281: 25425–25437.

11. RoikoMS, CarruthersVB (2009) New roles for perforins and proteases in apicomplexan egress. Cell Microbiol 11: 1444–1452.

12. RosenthalPJ (2004) Cysteine proteases of malaria parasites. Int J Parasitol 34: 1489–1499.

13. SturmA, AminoR, van de SandC, RegenT, RetzlaffS, et al. (2006) Manipulation of host hepatocytes by the malaria parasite for delivery into liver sinusoids. Science (New York, NY 313: 1287–1290.

14. HeusslerV, RennenbergA, StanwayR (2010) Host cell death induced by the egress of intracellular Plasmodium parasites. Apoptosis 15: 376–385.

15. AlyAS, MatuschewskiK (2005) A malarial cysteine protease is necessary for Plasmodium sporozoite egress from oocysts. J Exp Med 202: 225–230.

16. CoppiA, Pinzon-OrtizC, HutterC, SinnisP (2005) The Plasmodium circumsporozoite protein is proteolytically processed during cell invasion. J Exp Med 201: 27–33.

17. MonteiroAC, AbrahamsonM, LimaAP, Vannier-SantosMA, ScharfsteinJ (2001) Identification, characterization and localization of chagasin, a tight-binding cysteine protease inhibitor in Trypanosoma cruzi. Journal of cell science 114: 3933–3942.

18. SantosCC, CoombsGH, LimaAP, MottramJC (2007) Role of the Trypanosoma brucei natural cysteine peptidase inhibitor ICP in differentiation and virulence. Molecular microbiology 66: 991–1002.

19. BesteiroS, CoombsGH, MottramJC (2004) A potential role for ICP, a Leishmanial inhibitor of cysteine peptidases, in the interaction between host and parasite. Molecular microbiology 54: 1224–1236.

20. RiekenbergS, WitjesB, SaricM, BruchhausI, ScholzeH (2005) Identification of EhICP1, a chagasin-like cysteine protease inhibitor of Entamoeba histolytica. FEBS letters 579: 1573–1578.

21. LaCrueAN, SivaguruM, WalterMF, FidockDA, JamesAA, et al. (2006) A ubiquitous Plasmodium protein displays a unique surface labeling pattern in sporozoites. Mol Biochem Parasitol 148: 199–209.

22. PandeyKC, SinghN, Arastu-KapurS, BogyoM, RosenthalPJ (2006) Falstatin, a cysteine protease inhibitor of Plasmodium falciparum, facilitates erythrocyte invasion. PLoS pathogens 2: e117.

23. SandersonSJ, WestropGD, ScharfsteinJ, MottramJC, CoombsGH (2003) Functional conservation of a natural cysteine peptidase inhibitor in protozoan and bacterial pathogens. FEBS letters 542: 12–16.

24. Figueiredo da SilvaAA, de Carvalho VieiraL, KriegerMA, GoldenbergS, ZanchinNI, et al. (2007) Crystal structure of chagasin, the endogenous cysteine-protease inhibitor from Trypanosoma cruzi. Journal of structural biology 157: 416–423.

25. HansenG, HeitmannA, WittT, LiH, JiangH, et al. (2011) Structural basis for the regulation of cysteine-protease activity by a new class of protease inhibitors in Plasmodium. Structure 19: 919–929.

26. AparicioIM, ScharfsteinJ, LimaAP (2004) A new cruzipain-mediated pathway of human cell invasion by Trypanosoma cruzi requires trypomastigote membranes. Infection and immunity 72: 5892–5902.

27. SantosCC, ScharfsteinJ, LimaAP (2006) Role of chagasin-like inhibitors as endogenous regulators of cysteine proteases in parasitic protozoa. Parasitology research 99: 323–324.

28. PandeyKC, WangSX, SijwaliPS, LauAL, McKerrowJH, et al. (2005) The Plasmodium falciparum cysteine protease falcipain-2 captures its substrate, hemoglobin, via a unique motif. Proceedings of the National Academy of Sciences of the United States of America 102: 9138–9143.

29. RennenbergA, LehmannC, HeitmannA, WittT, HansenG, et al. (2010) Exoerythrocytic Plasmodium parasites secrete a cysteine protease inhibitor involved in sporozoite invasion and capable of blocking cell death of host hepatocytes. PLoS pathogens 6: e1000825.

30. PeiY, MillerJL, LindnerSE, VaughanAM, ToriiM, et al. (2013) Plasmodium yoelii inhibitor of cysteine proteases is exported to exomembrane structures and interacts with yoelipain-2 during asexual blood-stage development. Cell Microbiol 15: 1508–1526.

31. BoysenKE, MatuschewskiK (2013) Inhibitor of cysteine proteases is critical for motility and infectivity of Plasmodium sporozoites. mBio 4: e00874-00813.

32. CarvalhoTG, ThibergeS, SakamotoH, MenardR (2004) Conditional mutagenesis using site-specific recombination in Plasmodium berghei. Proceedings of the National Academy of Sciences of the United States of America 101: 14931–14936.

33. CombeA, GiovanniniD, CarvalhoTG, SpathS, BoissonB, et al. (2009) Clonal conditional mutagenesis in malaria parasites. Cell host & microbe 5: 386–396.

34. LacroixC, GiovanniniD, CombeA, BargieriDY, SpathS, et al. (2011) FLP/FRT-mediated conditional mutagenesis in pre-erythrocytic stages of Plasmodium berghei. Nature protocols 6: 1412–1428.

35. GiovanniniD, SpathS, LacroixC, PerazziA, BargieriD, et al. (2011) Independent roles of apical membrane antigen 1 and rhoptry neck proteins during host cell invasion by apicomplexa. Cell host & microbe 10: 591–602.

36. StanwayRR, GraeweS, RennenbergA, HelmS, HeusslerVT (2009) Highly efficient subcloning of rodent malaria parasites by injection of single merosomes or detached cells. Nature protocols 4: 1433–1439.

37. CoppiA, NatarajanR, PradelG, BennettBL, JamesER, et al. (2011) The malaria circumsporozoite protein has two functional domains, each with distinct roles as sporozoites journey from mosquito to mammalian host. J Exp Med 208: 341–356.

38. SultanAA, ThathyV, FrevertU, RobsonKJ, CrisantiA, et al. (1997) TRAP is necessary for gliding motility and infectivity of plasmodium sporozoites. Cell 90: 511–522.

39. EjigiriI, RaghebDR, PinoP, CoppiA, BennettBL, et al. (2012) Shedding of TRAP by a rhomboid protease from the malaria sporozoite surface is essential for gliding motility and sporozoite infectivity. PLoS pathogens 8: e1002725.

40. BaerK, KlotzC, KappeSH, SchniederT, FrevertU (2007) Release of hepatic Plasmodium yoelii merozoites into the pulmonary microvasculature. PLoS pathogens 3: e171.

41. MorahanBJ, SallmannGB, HuestisR, DubljevicV, WallerKL (2009) Plasmodium falciparum: genetic and immunogenic characterisation of the rhoptry neck protein PfRON4. Experimental parasitology 122: 280–288.

42. KappeS, BrudererT, GanttS, FujiokaH, NussenzweigV, et al. (1999) Conservation of a gliding motility and cell invasion machinery in Apicomplexan parasites. The Journal of cell biology 147: 937–944.

43. KappeSH, BuscagliaCA, BergmanLW, CoppensI, NussenzweigV (2004) Apicomplexan gliding motility and host cell invasion: overhauling the motor model. Trends in parasitology 20: 13–16.

44. MunterS, SabassB, Selhuber-UnkelC, KudryashevM, HeggeS, et al. (2009) Plasmodium sporozoite motility is modulated by the turnover of discrete adhesion sites. Cell host & microbe 6: 551–562.

45. HeggeS, MunterS, SteinbuchelM, HeissK, EngelU, et al. (2010) Multistep adhesion of Plasmodium sporozoites. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 24: 2222–2234.

46. SoldatiD, DubremetzJF, LebrunM (2001) Microneme proteins: structural and functional requirements to promote adhesion and invasion by the apicomplexan parasite Toxoplasma gondii. Int J Parasitol 31: 1293–1302.

47. ParussiniF, CoppensI, ShahPP, DiamondSL, CarruthersVB (2010) Cathepsin L occupies a vacuolar compartment and is a protein maturase within the endo/exocytic system of Toxoplasma gondii. Molecular microbiology 76: 1340–1357.

48. AgarwalS, SinghMK, GargS, ChitnisCE, SinghS (2013) Ca(2+) -mediated exocytosis of subtilisin-like protease 1: a key step in egress of Plasmodium falciparum merozoites. Cell Microbiol 15: 910–921.

49. Arastu-KapurS, PonderEL, FonovicUP, YeohS, YuanF, et al. (2008) Identification of proteases that regulate erythrocyte rupture by the malaria parasite Plasmodium falciparum. Nature chemical biology 4: 203–213.

50. ChildMA, EppC, BujardH, BlackmanMJ (2010) Regulated maturation of malaria merozoite surface protein-1 is essential for parasite growth. Molecular microbiology 78: 187–202.

51. LilburnTG, CaiH, ZhouZ, WangY (2011) Protease-associated cellular networks in malaria parasite Plasmodium falciparum. BMC genomics 12 Suppl 5: S9.

52. Schmidt-ChristensenA, SturmA, HorstmannS, HeusslerVT (2008) Expression and processing of Plasmodium berghei SERA3 during liver stages. Cell Microbiol 10: 1723–1734.

53. SinnisP, CoppiA (2007) A long and winding road: the Plasmodium sporozoite's journey in the mammalian host. Parasitology international 56: 171–178.

54. KangJM, JuHL, YuJR, SohnWM, NaBK (2012) Cryptostatin, a chagasin-family cysteine protease inhibitor of Cryptosporidium parvum. Parasitology 139: 1029–1037.

55. SantosCC, Sant'annaC, TerresA, Cunha-e-SilvaNL, ScharfsteinJ, et al. (2005) Chagasin, the endogenous cysteine-protease inhibitor of Trypanosoma cruzi, modulates parasite differentiation and invasion of mammalian cells. Journal of cell science 118: 901–915.

56. EngelJC, DoylePS, PalmerJ, HsiehI, BaintonDF, et al. (1998) Cysteine protease inhibitors alter Golgi complex ultrastructure and function in Trypanosoma cruzi. Journal of cell science 111: 597–606.

57. AminoR, ThibergeS, BlazquezS, BaldacciP, RenaudO, et al. (2007) Imaging malaria sporozoites in the dermis of the mammalian host. Nature protocols 2: 1705–1712.

58. JanseCJ, RamesarJ, WatersAP (2006) High-efficiency transfection and drug selection of genetically transformed blood stages of the rodent malaria parasite Plasmodium berghei. Nature protocols 1: 346–356.

59. StanwayRR, WittT, ZobiakB, AepfelbacherM, HeusslerVT (2009) GFP-targeting allows visualization of the apicoplast throughout the life cycle of live malaria parasites. Biology of the cell/under the auspices of the European Cell Biology Organization 101: 415–430 415 p following 430.

60. YoshidaN, NussenzweigRS, PotocnjakP, NussenzweigV, AikawaM (1980) Hybridoma produces protective antibodies directed against the sporozoite stage of malaria parasite. Science 207: 71–73.

61. EjigiriI, SinnisP (2009) Plasmodium sporozoite-host interactions from the dermis to the hepatocyte. Current opinion in microbiology 12: 401–407.

62. TsujiM, MatteiD, NussenzweigRS, EichingerD, ZavalaF (1994) Demonstration of heat-shock protein 70 in the sporozoite stage of malaria parasites. Parasitology research 80: 16–21.