Abstract
Intracellular leukoproliferative Theileria are unique as eukaryotic organisms that transform the immune cells of their ruminant host. Theileria utilize the uncontrolled proliferation for rapid multiplication and distribution into host daughter cells. The parasite distribution into the daughter cells is accompanied by a tight association with the host cell mitotic apparatus. Since the molecular basis for this interaction is largely unknown, we investigated the possible involvement of the immunodominant Theileria annulata surface protein, TaSP, in the attachment of the parasite to host cell microtubule network. Confocal microscopic analyses showed co-localization of the TaSP protein with alpha-tubulin and reciprocal immuno-co-precipitation experiments demonstrated an association of TaSP with alpha-tubulin in vivo. In addition, the partially expressed predicted extracellular domain of TaSP co-localized with the mitotic spindle of dividing cells and was co-immunoprecipitated with alpha-tubulin in transiently transfected Cos-7 cells devoid of other T. annulata expressed proteins. Pull-down studies showed that there is a direct interaction between TaSP and polymerized microtubules. Analysis of the interaction of TaSP and host microtubulin during host cell mitosis indicated that TaSP co-localizes and interacts with the spindle poles, the mitotic spindle apparatus and the mid-body. Moreover, TaSP was demonstrated to be localized to the microtubule organizing center and to physically interact with gamma-tubulin. These data support the notion that the TaSP—microtubule interaction may be playing a potential role in parasite distribution into daughter host cells and give rise to the speculation that TaSP may be involved in regulation of microtubule assembly in the host cell.
Similar content being viewed by others
References
Ahmed JS, Rothert M, Steuber S, Schein E (1989) In vitro proliferative and cytotoxic responses of PBL from Theileria annulata-immune cattle. Zentralbl Veterinarmed A 36:584–592
Bakheit MA, Endl E, Ahmed JS, Seitzer U (2006) Purification of macroschizonts of a Sudanese isolate of Theileria lestoquardi (T. lestoquardi [Atbara]). Ann N Y Acad Sci 1081:453–462
Baumgartner M, Tardieux I, Ohayon H, Gounon P, Langsley G (1999) The use of nocodazole in cell cycle analysis and parasite purification from Theileria parva-infected B cells. Microbes Infect 1:1181–1188
Baylis HA, Allsopp BA, Hall R, Carrington M (1993) Characterisation of a glutamine- and proline-rich protein (QP) from Theileria parva. Mol Biochem Parasitol 61:171–178
Biermann R, Schnittger L, Beyer D, Ahmed JS (2003) Initiation of translation and cellular localization of Theileria annulata casein kinase II alpha: implication for its role in host cell transformation. J Cell Physiol 196:444–453
Blom N, Sicheritz-Ponten T, Gupta R, Gammeltoft S, Brunak S (2004) Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence. Proteomics 4:1633–1649
Dobbelaere D, Heussler VT (1999) Transformation of leukocytes by Theileria parva and T. annulata. Annu Rev Microbiol 55:1–42
Dorée M, Hunt T (2002) From Cdc2 to Cdk1: when did the cell cycle kinase join its cyclin partner? J Cell Sci 115:2461–2464
Dschunkowsky E, Luhs J (1904) Die Piroplasmose der Rinder. Centralbl Bakteriol Parasitenk Infektion Hygiene 35:486–493
Ebel T, Middleton JF, Frisch A, Lipp J (1997) Characterization of a secretory type Theileria parva glutaredoxin homologue identified by a novel screening procedure. J Biol Chem 272:3042–3048
Fawcett D, Musoke A, Voigt W (1984) Interaction of sporozoites of Theileria parva with bovine lymphocytes in vitro. I. Early events after invasion. Tissue Cell 16:873–884
Frénal K, Soldati-Favre D (2009) Role of the parasite and host cytoskeleton in apicomplexa parasitism. Cell Host Microbe 5:602–611
Galley Y, Hagens G, Glaser I, Davis W, Eichhorn M, Dobbelaere D (1997) Jun NH2-terminal kinase is constitutively activated in T cells transformed by the intracellular parasite Theileria parva. Proc Natl Acad Sci USA 94:5119–5124
Gerber S (2009) Assoziation mit dem Wirtszell-Mikrotubulinetzwerk und zytoplasmatische Arretierung von p53: Untersuchungen zur Überlebensstrategie von Theileria annulata in transformierten Rinderleukozyten. Dissertation, University of Lübeck, Germany
Haller D, Mackiewicz M, Gerber S, Beyer D, Kullmann B, Schneider I, Ahmed JS, Seitzer U (2009) Cytoplasmic sequestration of p53 promotes survival in leukocytes transformed by Theileria
Heussler VT, Kuenzi P, Fraga F, Schwab RA, Hemmings BA, Dobbelaere DA (2001) The Akt/PKB pathway is constitutively activated in Theileria-transformed leucocytes, but does not directly control constitutive NF–kappaB activation. Cell Microbiol 3:537–550
Heussler VT, Rottenberg S, Schwab R, Kuenzi P, Fernandez PC, McKellar S, Shiels B, Chen ZJ, Orth K, Wallach D, Dobbelaer D (2002) Hijacking of host cell IKK signalosomes by the transforming parasite Theileria. Science 298:1033–1036
Hulliger L, Wilde KH, Brown CG, Turner L (1964) Mode of multiplication of Theileria in cultures of bovine lymphocytic cells. Nature 203:728–730
Irvin AD, Morrison WI (1987) Immunopathology, immunology and immunoprophylaxis of Theileria infections. In: Soulsby EL (ed) Protozoa, vol. III. Immune responses in parasitic infections: immunology, immunopathology and immunoprophylaxis. CRC Press, Boca Raton, pp 1–10
McHardy N, Wekesa LS, Hudson AT, Randall AW (1985) Antitheilerial activity of BW720C (buparvaquone): a comparison with parvaquone. Res Vet Sci 39:29–33
ole-MoiYoi OK (1995) Casein kinase II in theileriosis. Science 267:834–836
ole-MoiYoi OK, Sugimoto C, Conrad PA, Macklin MD (1992) Cloning and characterization of the casein kinase II alpha subunit gene from the lymphocyte-transforming intracellular protozoan parasite Theileria parva. Biochemistry 31:6193–61202
Schneider I, Haller D, Seitzer U, Beyer D, Ahmed JS (2004) Molecular genetic characterization and subcellular localization of a putative Theileria annulata membrane protein. Parasitol Res 94:405–415
Schneider I, Haller D, Kullmann B, Beyer D, Ahmed JS, Seitzer U (2007) Identification, molecular characterization and subcellular localization of a Theileria annulata parasite protein secreted into the host cell cytoplasm. Parasitol Res 101:1471–1482
Schnittger L, Katzer F, Biermann R, Shayan P, Boguslawski K, McKellar S, Beyer D, Shiels BR, Ahmed JS (2002) Characterization of a polymorphic Theileria annulata surface protein (TaSP) closely related to PIM of Theileria parva: implications for use in diagnostic tests and subunit vaccines. Mol Biochem Parasitol 120:247–256
Seitzer U, Schnittger L, Boguslawski K, Ahmed JS (2006) Investigation of MAP kinase activation in Theileria-infected cell lines. Ann N Y Acad Sci 1081:473–475
Seitzer U, Bakheit MA, Salih DE, Ali A, Haller D, Yin H, Schnittger L, Ahmed J (2007) From molecule to diagnostic tool: Theileria annulata surface protein TaSP. Parasitol Res 101(S2):S217–S223
Shayan P, Ahmed JS (1997) Theileria-mediated constitutive expression of the casein kinase II-alpha subunit in bovine lymphoblastoid cells. Parasitol Res 83:526–532
Shiels BR, McKellar S, Katzer F, Lyons K, Kinnaird J, Ward C, Wastling JM, Swan D (2004) A Theileria annulata DNA binding protein localized to the host cell nucleus alters the phenotype of a bovine macrophage cell line. Eukaryot Cell 3:495–505
Skilton RA, Bishop RP, Wells CW, Spooner PR, Gobright E, Nkonge C, Musoke AJ, Macklin M, Iams KP (1998) Cloning and characterization of a 150 kDa microsphere antigen of Theileria parva that is immunologically cross-reactive with the polymorphic immunodominant molecule (PIM). Parasitology 117:321–330
Stagg DA, Chasey D, Young AS, Morzaria SP, Dolan TT (1980) Synchronization of the division of Theileria macroschizonts and their mammalian host cells. Ann Trop Med Parasitol 74:263–265
Swan DG, Phillips K, Tait A, Shiels BR (1999) Evidence for localisation of a Theileria parasite AT hook DNA-binding protein to the nucleus of immortalised bovine host cells. Mol Biochem Parasitol 101:117–129
Swan DG, Stadler L, Okan E, Hoffs M, Katzer F, Kinnaird J, McKellar S, Shiels BR (2003) TashHN, a Theileria annulata encoded protein transported to the host nucleus displays an association with attenuation of parasite differentiation. Cell Microbiol 5:947–956
Tilney LG, Tilney MS, Shaw MK (1994) Theileria: strategy of infection and survival. In: Russel DG (ed) Baillières clinical infectious diseases: strategies for intracellular survival of microbes. Baillère Tindall, London, pp 335–350
Toye PG, Metzelaar MJ, Wijngaard PL, Nene V, Iams K, Roose J, Nyanjui JK, Gobright E, Musoke AJ, Clevers HC (1995) Characterization of the gene encoding the polymorphic immunodominant molecule, a neutralizing antigen of Theileria parva. J Immunol 155:1370–1381
Acknowledgements
We would like to thank Dr. Prim Singh for critical reading of the manuscript. This work was financed in part by EU-funded INCO-DEV projects ICTTD-3 (project number 510561) and ADDAV (project number ICA-1999-30151).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Seitzer, U., Gerber, S., Beyer, D. et al. Schizonts of Theileria annulata interact with the microtubuli network of their host cell via the membrane protein TaSP. Parasitol Res 106, 1085–1102 (2010). https://doi.org/10.1007/s00436-010-1747-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-010-1747-8