Belmont University Research Symposium (BURS)

Establishing a Biochemical System for the Purification of GST-Dbp5 to assess ATPase activity

Publication Date



Sciences and Mathematics, College of


Chemistry and Physics, Department of

BURS Faculty Advisor

Dr. Rachel E. Rigsby, PhD

Presentation Type

Poster Presentation


The export of mRNA out of the nucleus, the site of its production, and to the cytoplasm, where proteins are translated, is a crucial step for eukaryotic gene expression. This is aided by Mex67 binding to the transcript, which allows export through the nuclear pore complexes (NPCs), selective doorways embedded in the nuclear envelope. Once the transcript reaches the cytoplasm, a protein known as Dbp5, bound to ATP, Gle1, and Nup42 aids in the directionality of mRNA export by binding to the mRNA strand and removing Mex67. Following interaction with RNA, Dbp5 then hydrolyzes ATP so that it unbinds the mRNA, allowing for enzyme recycling. Therefore, Dbp5 ATPase activity is essential for mRNA export and cell viability. Factors that influence this activity can be accessed through biochemical purification and in vitro analysis. The goal of this work has been to enhance purification of recombinant Dbp5 for this analysis. Specifically, I have focused on optimizing induction of GST-Dbp5 in E. coli Rosetta cells. These efforts have yielded successful induction of Dbp5 at 16°C, and ongoing experiments are aimed at biochemical purification from induced cells. Future experiments will work towards establishing a biochemical system to analyze the ATPase activity of GST-Dbp5.

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