Processing of Toxoplasma ROP1 protein in nascent rhoptries

Secretion in the obligate intracellular parasite, Toxoplasma gondii, occurs through a number of regulated compartments. Among these are the apical organelles known as rhoptries which release their contents as part of the invasion process. We are interested in the processing, targeting and ultimate function of rhoptry proteins (and have focused our analyses on rhoptry protein 1 (ROP1). In this paper, we address the issue of processing: using a number of engineered forms of the ROP1 gene (introduced into a ROP1- background), we show that ROP1 is synthesized as a pre-pro-protein that is subject to proteolytic cleavages to remove the pre-sequence and the 'pro' region, at the N-terminus. Using brefeldin A (BFA) and reduced temperature we show that this processing occurs late in the secretory pathway of the parasite. Immunolocalization studies with epitope-tagged constructs indicate that processing is apparently occurring in the nascent rhoptries of dividing parasites. The results are discussed in the context of the targeting and possible function of the ROP1 protein.