Identifying Genetic Factors of Nuclear Pore Complex Assembly in Sacchyromyces cerevesiae

Abstract

The Nuclear Pore Complex (NPC), a 60-110 MDa protein complex embedded in the nuclear envelope, functions to transport all material across the nuclear membrane. While NPC structure is studied extensively, there is still much to learn about the assembly process. We are working to identify key factors to the assembly process by identifying the genetic mutation in the Saccharomyces cerevisiae NPC assembly mutant KRY141 through complementation testing. Fluorescence microscopy reveals the mislocalization of NPC proteins due to the failure to assemble NPCs. Since the single mutation confers both defects in NPC assembly (npa) and temperature sensitivity (ts) phenotypes, we use temperature sensitivity as a measure for complementation of the mutation. Previous work mapped the NPC assembly mutation near the MAT locus on chromosome III. Since then, genes in the region have been isolated one at a time to attempt to rescue the mutant phenotype. In this way, we have ruled out 4 potential genes: MAK32, PMP1, YCR024C-B, and YCR025C. Identification of this gene required in the assembly of the NPC would provide key information on an essential biological process and provide insight to NPC assembly related diseases associated with aging and cancers.