Yeast Two-Hybrid (Y2H) to Test Dbp5-Gle1 Interaction Dependence on FG domains and Mex67

Publication Date

Spring 2026

Presentation Length

Poster/Gallery presentation

College

College of Sciences & Mathematics

Department

Biology, Department of

Student Level

Undergraduate

Faculty Mentor

Rebecca Adams, PhD

Presentation Type

Poster

Summary

For proteins to be translated in the cytoplasm of a eukaryotic cell, mRNA must first be exported out of the nucleus, where it is made, and through nuclear pore complexes (NPC), selective doorways embedded in the nuclear envelope. In yeast cells, Mex67 binds to and guides mature mRNA out the nucleus through the NPC. At the cytoplasmic side of the NPC, the nucleoporin protein Nup42 facilitates the removal of Mex67 from the mRNA transcript by regulating the enzymatic activity of Dpb5 and its binding partner Gle1, which bind to the transcript to release Mex67. This enables unidirectional transport of mRNA across the NPC. Specifically, Gle1 binds to Dpb5 so Dpb5 can then remove Mex67 from the mRNA transcript, and Gle1 also binds to Nup42 on its C-terminal domain (CTD). The N-terminal domain (NTD) of Nup42 is an FG domain that binds to Mex67 and believed to be required for the Gle1-Dpb5 interaction to occur. The goal in this experiment is to test two different aspects of the Gle1-Dpb5 interaction with Nup42. We hypothesize that the FG domain of Nup42 is required for Gle1 and Dpb5 to interact, and we sought to explore whether the FG domain of other NPC proteins can also serve in this role. To test this, we performed yeast two-hybrid (Y2H) assays with different Gle1 and Dpb5 Y2H constructs to observe the protein interactions in the absence of Nup42. Along with the Gle1 and Dpb5 Y2H constructs, variations of Nup42 swapped with different FG domains was used to test FG binding of Gle1.

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