Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
권호 표지

Nucleolar translocalization of GRA10 of Toxoplasma gondii transfectionally expressed in HeLa cells

  • Ahn, Hye-Jin (Department of Parasitology and the Catholic Institute of Parasitic Diseases, College of Medicine, Catholic University of Korea) ;
  • Kim, Sehra (Department of Parasitology and the Catholic Institute of Parasitic Diseases, College of Medicine, Catholic University of Korea) ;
  • Nam, Ho-Woo (Department of Parasitology and the Catholic Institute of Parasitic Diseases, College of Medicine, Catholic University of Korea)
  • Published : 2007.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

Toxoplasma gondii GRA10 expressed as a GFP-GRA10 fusion protein in HeLa cells moved to the nucleoli within the nucleus rapidly and entirely. GRA10 was concentrated specifically in the dense fibrillar component of the nucleolus morphologically by the overlap of GFP-GRA10 transfection image with IFA images by monoclonal antibodies against GRA10 (Tg378), B23 (nucleophosmin) and C23 (nucleolin). The nucleolar translocalization of GRA10 was caused by a putative nucleolar localizing sequence (NoLS) of GRA10. Interaction of GRA10 with TATA-binding protein associated factor 1B (TAF1B) in the yeast two-hybrid technique was confirmed by GST pull-down assay and immunoprecipitation assay. GRA10 and TAF1B were also co-localized in the nucleolus after co-transfection. The nucleolar condensation of GRA10 was affected by actinomycin D. Expressed GFP-GRA10 was evenly distributed over the nucleoplasm and the nucleolar locations remained as hollows in the nucleoplasm under a low dose of actinomycin D. Nucleolar localizing and interacting of GRA10 with TAF1B suggested the participation of GRA10 in rRNA synthesis of host cells to favor the parasitism of T. gondii.

Keywords

References

  1. Ahn HJ, Kim S, Nam HW (2005) Host cell binding of GRA10, a novel, constitutively secreted dense granular protein from Toxoplasma gondii. Biochem Biophys Res Comm 331: 614-620 https://doi.org/10.1016/j.bbrc.2005.03.218
  2. Ahn HJ, Kim S, Kim HE, Nam HW (2006) Interactions between secreted GRA proteins and host cell proteins across the parasitophorous vacuolar membrane in the parasitism of Toxoplasma gondii. Korean J Parasitol 44: 303-312 https://doi.org/10.3347/kjp.2006.44.4.303
  3. Chen D, Huang S (2001) Nucleolar components involved in ribosome biogenesis cycle between the nucleolus and nucleoplasm in interphase cells. J Cell Biol 153: 169-176 https://doi.org/10.1083/jcb.153.1.169
  4. Coleman NA, Peeples ME (1993) The matrix protein of Newcastle disease virus localizes to the nucleus via a bipartite nuclear localization signal. Virology 195: 596- 607 https://doi.org/10.1006/viro.1993.1411
  5. Dang CV, Lee WM (1988) Identification of the human c-myc protein nuclear translocation signal. Mol Cell Biol 8: 4048-4054 https://doi.org/10.1128/MCB.8.10.4048
  6. Dang CV, Lee WM (1989) Nuclear and nucleolar targeting sequences of c-erb-A, c-myb, N-myb, p53, HSP70, and HIV tat proteins. J Biol Chem 264: 18019-18023
  7. Dundr M, Leno GH, Hammarskjold ML, Rekosh D, Helga-Maria C, Olson MOJ (1995) The roles of nucleolar structure and function in the subcellular location of the HIV-1 Rev protein. J Cell Sci 108: 2811-2823
  8. Fan Q, An L, Cui L (2004) Plasmodium falciparum histone acetyltransferase, a yeast GCN5 homologue involved in chromatin remodeling. Eukaryotic Cell 3: 264-276 https://doi.org/10.1128/EC.3.2.264-276.2004
  9. Favre D, Studer E, Michel MR (1994) Two nucleolar targeting signals present in the N-terminal part of Semliki Forest virus capsid protein. Arch Virol 137: 149-155 https://doi.org/10.1007/BF01311181
  10. Friedrich JK, Panov KI, Cabart P, Russell J, Zomerdijk JCBM (2005) TBP-TAF complex SL1 directs RNA polymerase I pre-initiation complex formation and stabilizes upstream binding factor at the rDNA promoter. J Biol Chem 280: 29551-29558 https://doi.org/10.1074/jbc.M501595200
  11. Gilbert LA, Ravindran S, Turetzky JM, Boothroyd JC, Bradley PJ (2007) Toxoplasma gondii targets a protein phosphatase 2C to the nucleus of infected host cells. Eukaryotic Cell 6: 73-83 https://doi.org/10.1128/EC.00309-06
  12. Grummt I (2003) Life on a planet of its own: regulation of RNA polymerase I transcription in the nucleus. Genes Develop 17: 1691-1702
  13. Grummt I, Pikaard CS (2003) Epigenetic silencing of RNA polymerase I transcription. Nat Rev Mol Cell Biol 4: 641- 649 https://doi.org/10.1038/nrm1171
  14. Gubbels MJ, Striepen B, Shastri N, Turkoz M, Robey EA (2005) Class I major histocompatibility complex presentation of antigens that escape from the parasitophorous vacuole of Toxoplasma gondii. Infect Immun 73: 703-711 https://doi.org/10.1128/IAI.73.2.703-711.2005
  15. Henderson JE, Amizuka N, Warshasky H, Biasotto D, Lanske BMK, Goltzman D, Karaplis AC (1995) Nucleolar localization of parathyroid hormone-related peptide enhances survival of chondrocytes under conditions that promote apoptotic cell death. Mol Cell Biol 15: 4064-4075 https://doi.org/10.1128/MCB.15.8.4064
  16. Hernandez-Verdun D (2006) The nucleolus: a model for the organization of nuclear functions. Histochem Cell Biol 126: 135-148 https://doi.org/10.1007/s00418-006-0212-3
  17. Hiscox JA (2002) The nucleolus: gateway to viral infection? Arch Virol 147: 1077-1089 https://doi.org/10.1007/s00705-001-0792-0
  18. Hoeck J, Woisetschlager M (2001) Activation of Eotaxin- 3/CCL26 gene expression in human dermal fibroblasts is mediated by STAT6. J Immunol 167: 3216-3222 https://doi.org/10.4049/jimmunol.167.6.3216
  19. Kiefer P, Dickson C (1995) Nucleolar association of fibroblast growth factor 3 via specific sequence motifs has inhibitory effects on cell growth. Mol Cell Biol 15: 4364- 4374 https://doi.org/10.1128/MCB.15.8.4364
  20. Louvet E, Junera HR, Le Panse S, Hernandez-Verdun D (2005) Dynamics and compartmentation of the nucleolar processing machinery. Exp Cell Res 304: 457-470 https://doi.org/10.1016/j.yexcr.2004.11.018
  21. Magno RC, Straker LC, de Souza W, Attias M (2005) Interrelations between the parasitophorous vacuole of Toxoplasma gondii and host cell organelles. Microsc Microanal 11: 166-174
  22. Malim MH, Bohnlein S, Hauber J, Cullen BR (1989) Functional dissection of the HIV-1 Rev trans-activator. Derivation of a trans-dominant repressor of Rev function. Cell 58: 205-214 https://doi.org/10.1016/0092-8674(89)90416-9
  23. Mercier C, Adjogble KDZ, Daubener W, Delauw MFC (2005) Dense granules: Are they key organelles to help understand the parasitophorous vacuole of all Apicomplexa parasites? Int J Parasitol 35: 829-849 https://doi.org/10.1016/j.ijpara.2005.03.011
  24. Ngo HM, Hoppe HC, Joiner JA (2000) Differential sorting and post-secretory targeting of proteins in parasitic invasion. Trends Cell Biol 10: 67-72 https://doi.org/10.1016/S0962-8924(99)01698-0
  25. Phair RD, Misteli T (2000) High mobility of proteins in the mammalian cell nucleus. Nature 404: 604-609 https://doi.org/10.1038/35007077
  26. Saharinen P, Takaluoma K, Silvennoinen O (2000) Regulation of the Jak2 tyrosine kinase by its pseudokinase domain. Mol Cell Biol 20: 3387-3395 https://doi.org/10.1128/MCB.20.10.3387-3395.2000
  27. Siomi H, Shida H, Nam SH, Nosaka T, Maki M, Hatanaka M (1988) Sequence requirements for nucleolar localization of human T cell leukemia virus type I pX protein, which regulates viral RNA processing. Cell 55: 197-209 https://doi.org/10.1016/0092-8674(88)90043-8
  28. Templeton TJ, Deitsch KW (2005) Targeting malaria parasite proteins to the erythrocyte. Trends Parasitol 21: 399-402 https://doi.org/10.1016/j.pt.2005.07.006
  29. Thiry M, Lafontaine DLJ (2005) Birth of a nucleolus: the evolution of nucleolar compartments. Trend Cell Biol 15: 194-199 https://doi.org/10.1016/j.tcb.2005.02.007
  30. Tilley L, McFadden G, Cowman A, Klonis N (2007) Illuminating Plasmodium falciparum-infected red blood cells. Trends Parasitol 23: 268-277 https://doi.org/10.1016/j.pt.2007.04.001
  31. Tonkin CJ, Pearce JA, McFadden GI, Cowman AF (2006) Protein targeting to destinations of the secretory pathway in the malaria parasite Plasmodium falciparum. Cur Opinion Microbiol 9: 381-387
  32. Valdez BC, Perlaky L, Henning D, Saijo Y, Chan PK, Busch H (1994) Identification of the nuclear and nucleolar localization signals of the protein p120. J Biol Chem 269: 23776-23783
  33. Zhao LJ, Padmanabhan R (1988) Nuclear transport of adenovirus DNA polymerase is facilitated by interaction with preterminal protein. Cell 55: 1005-1015 https://doi.org/10.1016/0092-8674(88)90245-0