The yeast prion [PSI+] is the misfolded version of the translation termination protein Sup35p. The study of [PSI+] has greatly enhanced our understanding of how prions propagate; yet the cellular mechanisms underlying how prions form are not well understood. It has been proposed that the initial misfolding and assembly of Sup35p aggregates accumulate at a site called IPOD, Insoluble Protein Deposit, which is tethered to the vacuole. To uncover other cellular pathways that may be involved in prion formation, we previously performed a small scale screen of deletion mutants. We identified several gene deletions that impaired early stages of prion formation and others that that affected later stages. Since most of the deletions possessed fragmented vacuoles, we originally hypothesized that vacuole integrity and the maintenance of its tethered component IPOD is essential for prion formation. Our current data indicate that vacuole fragmentation and prion formation are independent of each other. In fact, introduction of a centromeric plasmid alone was sufficient to cause fragmentation but not impact prion formation. However, it appears that gene deletions that impair early stages of prion formation did have altered localization of IPOD. This alteration in IPOD is not seen in wild type strains or gene deletions that affect later stages. Together our data suggest that although vacuole fragmentation does not affect prion formation, its tethered component IPOD likely plays an important role in the early stages of prion formation. Efforts to further characterize IPOD as it relates to prion formation will be presented. .