Identification and Characterization of a Mutated Gene Affecting NPC assembly

Abstract

The nuclear pore complex (NPC) is a gateway that controls what is exchanged across the nuclear envelope. Therefore, when something affects its assembly, the cell will die. The laboratory has a collection of Saccharomyces cerevisiae mutant strains with NPC assembly defects, but where these defects are located and how they are related to NPC assembly remains unknown. The goal of this project is to identify the mutated gene that is causing assembly defects within one of the strains. Fluorescent microscopy was used to confirm the presence of a NPC assembly defect, and the growth rate of the mutant strain was measured at the permissive and nonpermissive temperature. Additionally, a genomic library was used to compliment the temperature sensitive defect as well as NPC assembly defect. Fluorescent microscopy confirmed the presence of an assembly defect. It was additionally found that the growth rate of the mutant was significantly slower at the nonpermissive temperature. Upon completing all transformations, none of the plasmids in the genomic library rescued the mutant. With information still missing by the mechanism of which NPC’s are formed, identifying and characterizing genes that affect NPC assembly can lead to more information about it.