Belmont University Research Symposium (BURS)

Publication Date

Spring 2024

College

Sciences and Mathematics, College of

Department

Biology, Department of

Presentation Type

Poster Presentation

Abstract

Assessing Rescue of Dbp5R369G by Mex67 Overexpression.

Yaneli Basilio-Rodriguez, Rebecca Adams, PhD

In order for proteins to be translated in eukaryotic cells, mature mRNA that is generated in the nucleus must travel through the nuclear pore complex (NPC), doorways embedded in the nuclear envelope, to reach ribosomes in the cytoplasm. This occurs as the transcript is bound by the export protein Mex67. Once out of the nucleus and in the cytoplasm, Mex67 is removed by protein Dbp5, which provides directionality to transport, as Mex67 is required for transport across the NPC. The transcript is also bound by additional proteins in the nucleus, and these remain associated with the mRNA even after export. The goal of this project is to uncover how Dbp5 selectively removes Mex67 from the mRNA. Previous studies have generated a mutation in S. cerevisiae Dbp5 (Dbp5R369G) that results in a protein that cannot bind to RNA, and which results in a dominant negative growth defect when expressed. We hypothesize that this growth defect is due to loss of Mex67 removal from the mRNA, and lack of recycling of Mex67 to permit additional rounds of mRNA export. Therefore, we anticipate that overexpression of Mex67 will restore viability in cells in the presence of Dbp5R369G. My work has been aimed at generating a Mex67 overexpression plasmid by first amplifying Mex67, and then cloning this into a Gal-responsive overexpression vector. Following the successful cloning of this plasmid, I will next transform this plasmid into a yeast strain that has a Gal- Dbp5R369G plasmid.

I anticipate that cell viability should be restored in the cells when Mex67 is expressed, and these results should provide information about how Dbp5 removes Mex67 from the transcript.

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