A HT/PEXEL motif in Toxoplasma dense granule proteins is a signal for protein cleavage but not export into the host cell.
Apicomplexan parasites, such as Toxoplasma gondii and Plasmodium, secrete proteins for attachment, invasion and modulation of their host cells. The host targeting (HT), also known as the Plasmodium export element (PEXEL), directs Plasmodium proteins into erythrocytes to remodel the host cell and establish infection. Bioinformatic analysis of Toxoplasma revealed a HT/PEXEL-like motif at the N-terminus of several hypothetical unknown and dense granule proteins. Hemagglutinin-tagged versions of these uncharacterized proteins show co-localization with dense granule proteins found on the parasitophorous vacuole membrane (PVM). In contrast to Plasmodium, these Toxoplasma HT/PEXEL containing proteins are not exported into the host cell. Site directed mutagenesis of the Toxoplasma HT/PEXEL motif, RxLxD/E, shows that the arginine and leucine residues are permissible for protein cleavage. Mutations within the HT/PEXEL motif that prevent protein cleavage still allow for targeting to the PV but the proteins have a reduced association with the PVM. Addition of a Myc tag before and after the cleavage site shows that processed HT/PEXEL protein has increased PVM association. These findings suggest that while Toxoplasma and Plasmodium share similar HT/PEXEL motifs, Toxoplasma HT/PEXEL containing proteins interact with but do not cross the PVM.
Department of Medical Microbiology & Immunology, University of Wisconsin-Madison, Madison, WI, USA.
Chia-Hung Christine
CH
Hsiao
N
N
Luisa Hiller
Kasturi
K
Haldar
Laura J
LJ
Knoll
D000465:Algorithms; D020816:Amino Acid Motifs; D000818:Animals; D000953:Antigens, Protozoan; D019295:Computational Biology; D003902:Detergents / Q000494:pharmacology; D005347:Fibroblasts / Q000469:parasitology; D006388:Hemagglutinins / Q000737:chemistry; D006801:Humans; D008856:Microscopy, Fluorescence; D016297:Mutagenesis, Site-Directed; D017830:Octoxynol; D010957:Plasmids / Q000378:metabolism; D011092:Polyethylene Glycols / Q000494:pharmacology; D011485:Protein Binding; D020033:Protein Isoforms / Q000737:chemistry; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D016271:Proto-Oncogene Proteins c-myc / Q000737:chemistry; D015800:Protozoan Proteins / Q000737:chemistry*; D014122:Toxoplasma / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism
D000953:Antigens, Protozoan; D003902:Detergents; D006388:Hemagglutinins; D020033:Protein Isoforms; D016271:Proto-Oncogene Proteins c-myc; D015800:Protozoan Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40