The C Isoform of Dictyostelium Tetraspanins Localizes to the Contractile Vacuole and Contributes to Resistance against Osmotic Stress.

摘要: Tetraspanins (Tsps) are membrane proteins that widely expressed in eukaryotic organisms. Only recently, Tsps have started to acquire relevance as potential new drug targets they contribute, via protein-protein interactions, numerous pathophysiological processes including infectious diseases and cancer. However, due a high number of isoforms functional redundancy, knowledge on specific functions most is still scarce. We set out characterize five previously annotated Tsps, TspA-E, from Dictyostelium discoideum, model for studying human orthologues. Using reverse transcriptase PCRs, we found mRNAs TspA-E the multicellular slug stage, whereas vegetative cells only TspA, TspC and, lesser extent, TspD. raised antibodies against TspD detected endogenous well heterologously TspA by Western blot. N-deglycosylation assays mutational analyses showed glycosylation vivo. GFP-tagged co-localized with proton pump contractile vacuole network. Deletion strains exibited unaltered growth, adhesion, random motility development. Yet, tspC− defect coping hypo-osmotic stress, accumulation vacuoles, but heterologous expression rescued their phenotype. In conclusion, our data fill gap research open up possibility vacuoles e.g. Trypanosoma may one day constitute valuable target treating sleeping sickness, threatening tropical diseases.