Kelly Styles - 1507612 The function of the parasitophorous vacuole in Toxoplasma gondii The apicomplexan parasite Toxoplasma gondii

is an intracellular obligate parasite whose definitve host is the domesticated cat. T. gondii forms a unique parasitophorous vacuole upon entering an appropriate host cell in a mammalian intermediate host. The role of this parasitophorous vacuole is to provide a stable environment for replication. It does this in several ways; it forms during host cell invasion, forming a distinct membrane; it provides the parasites with a stable environment; it prevents the host cell from eliminating the parasites; and it facilitates the uptake of nutrients from the host cell. This parasitophorous vacuole is one of the factors that makes T. gondii one of the most successful human parasites. The parasitophorous vacuole forms when T. gondii enters a mammalian cell [1]. It uses the host cells plasma membrane to form the interface between the host cell cytosol and the parasitophorous vacuole (PV), the parasitophorous vacuole membrane (PVM) [1]. The PVM ensures that the host cell does not lyse during cell invasion, otherwise the parasite would not gain access to the nutrients inside the host cell [2]. The life stage of T. gondii that penetrates a host cell is the tachyozite [3]. The tachyozites, upon reaching the host plasma membrane, discharge the contents of the rhoptries and dense granules; organelles containing proteins and perhaps phospholipids, for growth of the PVM [3, 4]. An experiment was designed with the aim of determining the origins of the phospholipids from the PV; by measuring the host cell electrical resistance using patch clamps [2]. The study found that at least 80% of the PVM phospholipids came from the host plasma membrane [2]. The lipids appear to flow from the host cells plasma membrane to the PVM through an exclusion zone called the moving junction; in which host proteins are excluded [3]. Therefore, most of the proteins in the PVM are of parasitic origin. The PVM protects the PV from fusing with other membranous structures within the host cell [1]. This is important in maintaining a stable environment for parasite growth. Fusion of the PV with a lysosome; a membrane bound region with a low pH, would acidify the PV and kill the T. gondii parasites growing inside [5]. The PVM is non-fusogenic because it lacks certain receptor proteins that mediate membrane to membrane fusion [6]. It is thought that these host cell membrane receptor proteins have a tail of protein extending into the cytoplasm, and are excluded as they move through the moving junction of the nascent PVM [6]. This has implications for the ability of the host cell to remove the parasites. Normally, cell membrane receptors would signal to the immune system that the PV is of foreign origin; and infected cells would undergo autophagy, or programmed cell death in an attempt to halt the replication and spread of the parasite [7]. A successful parasite, like T. gondii, is able to prevent autophagy so it is continually supplied with nutrients for growth, and to provide time for further development into the bradyzoite lifestage of T. gondii [8]. The parasites are able to hide from the host cells immune system because the PVM lacks the receptor protein CD40 [5]. CD40 induces autophagy; the direction of organelles to the lysosomes, where the T. gondii parasites will be killed [5]. Preventing autophagy is therefore a key role of the PV in protecting T. gondii whilst in the intracellular environment. However, the distinct molecular nature of the PVM has some drawbacks. The non-fusogenic nature of the PVM means that nutrients contained in host cell vesicles from the secretory pathway cannot be obtained through fusion [1]. Instead T. gondii employs several mechanisms for obtaining nutrients. The PVM itself is permeable to molecules of a certain 1

Kelly Styles - 1507612 size, allowing exchange of molecules that range in size from 1300 to 1900 Daltons [9]. This is possible through the use of parasitic membrane spanning proteins which act as pores through which molecules can travel [9]. It is thought that these proteins originated from the rhoptries and dense granules [9]. Therefore, the parasites can obtain amino acids; monosaccharides; and nucleotides, which should be able to freely diffuse through the PVM [9]. T. gondii needs a different mechanism to obtain larger macronutrients. The endoplasmic reticulum and mitochondria are drawn into close proximity with the PV. This is thought to enable the transfer of nutrients between these organelles and the PV [1]. There exist junctions between the endoplasmic reticulum and the PV, suggesting that there is transfer of some material between the two compartments; including phospholipids [10]. Cholesterols essential for T. gondii growth are also accumulated from the hosts extracellular environment to the PVM by the low density lipoprotein pathway [11]. The mitochondria may be a source of lipoic acids; another vital lipid that is needed in the Tricarboxylic Acid Cycle [10]. This ensures that the parasite has the necessary nutrients for metabolism, and therefore growth. T. gondii is a very successful parasite, whose parasitophorous vacuole enables it to survive and replicate in the intracellular environment of the mammalian cell. The parasitophorous vacuole membrane is formed in such a way that host membrane phospholipids are incorporated into the growing PVM, but host membrane proteins with cytoplasmic tails are excluded. This produces the non-fusogenic nature of the PVM, protecting it from fusing with a lysosome which would kill the parasites inside the PV. The PVM also lacks the receptor protein CD40 which hides it from the hosts immune system, effectively stopping autophagy in the infected cells. The parasites obtain nutrition from the host through protein channels in the PVM which allow molecules of a certain size to pass through. There is also thought to be transfer of lipids from the endoplasmic reticulum and the mitochondrion which are closely associated with the PV. The role of the PV is therefore essential for the successful impregnation and replication of the parasites inside a host cell.

References 1. 2. Lalibert, J., and & Carruthers, V. (2008). Host cell manipulation by the human pathogen Toxoplasma gondii. Cellular and Molecular Life Sciences 65, 1900-1915. Suss-Toby, E., Zimmerberg, J., and Ward, G.E. (1996). Toxoplasma invasion: the parasitophorous vacuole is formed from host cell plasma membrane and pinches off via a fission pore. Proceedings of the National Academy of Sciences 93, 8413-8418. Lingelbach, K., and Joiner, K. (1998). The parasitophorous vacuole membrane surrounding Plasmodium and Toxoplasma: an unusual compartment in infected cells. J Cell Sci 111, 14671475. Con, J.M.B., Dubremetz, J.-F., Mercereau-Puijalon, O., and Joiner, K.A. (1994). The Toxoplasma gondii Rhoptry Protein ROP 2 Is Inserted into the Parasitophorous Vacuole Membrane, Surrounding the Intracellular Parasite, and Is Exposed to the Host Cell Cytoplasm. The Journal of Cell Biology 127, 947-961. Subauste, C.S. (2009). CD40, autophagy and Toxoplasma gondii. Memrias do Instituto Oswaldo Cruz 104, 267-272.

3.

4.

5.

Kelly Styles - 1507612 6. Joiner, K.A., Fuhrman, S.A., Miettinen, H.M., Kasper, L.H., and Mellman, I. (1990). Toxoplasma gondii: fusion competence of parasitophorous vacuoles in Fc receptortransfected fibroblasts. Science 249, pp641(646). Hippe, D., Weber, A., Zhou, L., Chang, D.C., Hacker, G., and Luder, C.G.K. (2009). Toxoplasma gondii infection confers resistance against BimS-induced apoptosis by preventing the activation and mitochondrial targeting of pro-apoptotic Bax. J Cell Sci 122, 3511-3521. Boyle, J.P., and Radke, J.R. (2009). A history of studies that examine the interactions of Toxoplasma with its host cell: Emphasis on in vitro models. International Journal for Parasitology 39, 903-914. Schwab, J.C., Beckers, C.J., and Joiner, K.A. (1994). The parasitophorous vacuole membrane surrounding intracellular Toxoplasma gondii functions as a molecular sieve. Proceedings of the National Academy of Sciences 91, 509-513. Sinai, A.P., and Joiner, K.A. (2001). The Toxoplasma gondii Protein ROP2 Mediates Host Organelle Association with the Parasitophorous Vacuole Membrane. The Journal of Cell Biology 154, 95-108. Coppens, I., Sinai, A.P., and Joiner, K.A. (2000). Toxoplasma gondii Exploits Host Low-Density Lipoprotein Receptor-Mediated Endocytosis for Cholesterol Acquisition. The Journal of Cell Biology 149, 167-180.

7.

8.

9.

10.

11.