Belmont University Research Symposium (BURS)

Publication Date

2024

College

Sciences and Mathematics, College of

Department

Biology, Department of

BURS Faculty Advisor

Rebecca Adams

Presentation Type

Poster Presentation

Abstract

In order for proteins to be made in eukaryotic cells, mRNA must first be exported from their site of production in the nucleus through the nuclear pore complex (NPC) to reach the cytoplasm. To cross through the NPC, mRNA binds to a transport protein called Mex67 which is able to bind NPC proteins via their FG domains to allow passage of the mRNA. Once in the cytoplasm, Mex67 is removed by Dbp5, and without Mex67 bound, the transcript cannot re-enter the nucleus, ensuring the directionality of mRNA export. The function of Dbp5 is regulated by its interaction with the protein Gle1, and the Dbp5-Gle1 interaction is dependent on the NPC protein Nup42. The N-terminus of Nup42 is an FG domain that binds to Mex67, and the Nup42 C-terminal domain (CTD) binds to Gle1. Previous experiments have demonstrated that the Nup42 FG domain is required for the Gle1-Dbp5 interaction, and we hypothesize that this is due to the interaction of this domain with Mex67. To test this hypothesis, we have performed a series of yeast two-hybrid (Y2H) experiments in S. cerevisiae to assess which regions of Nup42 permit the Dbp5-Gle1 interaction, whether other FG domains can compensate for the Nup42 FG domain, and whether fusing Mex67 to the Nup42 CTD will permit Gle1-Dbp5 binding. Importantly, through these studies, we discovered that in fact, Gle1 and Dbp5 do display a Y2H interaction in the absence of Nup42 when the construct includes the full length of Gle1. Therefore, future experiments will be used to explore the mechanism of Nup42 function with truncated Gle1.

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