Belmont University Research Symposium (BURS)

Publication Date

2024

College

Sciences and Mathematics, College of

Department

Biology, Department of

BURS Faculty Advisor

Dr. Rebecca Adams

Presentation Type

Poster Presentation

Abstract

The export of mRNA out of the nucleus, where it is generated, and into the cytoplasm where it is translated, is a crucial step for eukaryotic gene expression. The export of mRNA transcripts is aided by Mex67, which allows export through the selective nuclear pore complex (NPC) doorways in the nuclear envelope. Once out of the nucleus, a protein known as Dbp5, bound to ATP, Gle1, and Nup42 aids in the directionality of mRNA export by removing Mex67 from the mRNA strand. Following interaction with RNA, Dbp5 then hydrolyzes ATP so that it unbinds the mRNA, allowing for enzyme recycling. The goal of this study is to explore the ATPase activity of Saccharomyces cerevisiae Dbp5 in vitro with purified recombinant protein. However, we have not had a system for protein purification established at Belmont, and my efforts have been aimed at optimizing purification of Dbp5. In previous work, I enhanced the bacterial induction of Dbp5 by using the Rosetta strain of the E. coli. Having established a system for expression, my recent goal has been to purify GST-Dpb5 from cells via affinity chromatography. My work has allowed for the successful purification of GST-Dbp5 in high yield. Future experiments will assess the ATPase activity of this purified protein so that subsequent experiments can test the effect of Nup42 and Gle1 on this activity.

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