Belmont University Research Symposium (BURS)

Testing if a Gle1-Mex67-Mtr2 fusion bypasses need for Nup42 in binding Dpb5.

Publication Date

2024

College

Sciences and Mathematics, College of

Department

Biology, Department of

BURS Faculty Advisor

Rebecca Adams

Presentation Type

Poster Presentation

Abstract

An essential part of the protein production process is the export of the mRNA transcript out of the nucleus into the cytoplasm. There are several highly conversed proteins that function in this process of mRNA export, and this study uses the budding yeast S. cerevisiae as a model for studying this process. Two of these proteins include Mex67-Mtr2, a dimer which permits mRNA export by ferrying the transcript through the nuclear pore complex (NPC), doorways embedded in the nuclear envelope. The additional proteins, Dpb5 and Gle1, interact to remove the Mex67-Mtr2 protein dimer from the transcript after it exits the nucleus. This process ensures that the mRNA transcript will not go back into the nucleus so that it may be translated. The NPC protein, Nup42, aids in mRNA export through interaction with Mex67-Mtr2 via its amino-terminal FG domain and interaction with Gle1 at its carboxy-terminal domain. Previous evidence has suggested that the Nup42 FG domain is required for Dbp5-Gle1 interaction as this interaction is lost in a Nup42 deletion mutant. We hypothesize that Nup42 serves as a bridge between Gle1 and Mex67-Mtr2 and that this function is needed for Gle1 to bind to Dpb5. To test this hypothesis, our goal was to generate a fusion protein between Gle1, Mex67, and Mtr2, to test if this fusion will permit Gle1-Dpb5 interaction in the absence of Nup42 using a yeast two-hybrid (Y2H) assay. the gene for Mtr2 was cloned into the Y2H GBD-Mex67-Gle1 plasmid that was already available. This plasmid will then be transformed into a Nup42 deletion strain that has the Y2H GAD-Dpb5 plasmid. The resulting strains will then be plated onto -His media where growth indicates a positive interaction between the two proteins. Growth of this strain will provide insight into the mechanistic role of Nup42 during mRNA export.

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