Science University Research Symposium (SURS)
Publication Date
2022
College
Sciences and Mathematics, College of
Department
Chemistry and Physics, Department of
SURS Faculty Advisor
Rachel E. Rigsby, PhD
Presentation Type
Poster Presentation
Abstract
The export of mRNA out of the nucleus is a crucial step for eukaryotic gene expression. The export of mRNA transcripts is aided by Mex67, which allows export through the nuclear pore complex 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 helping remove Mex67 from the mRNA strand. Following interaction with RNA, Dbp5 then hydrolyzes ATP so that it unbinds the mRNA, allowing for enzyme recycling. Previous efforts worked towards the purification of Dbp5, but the attempts were unsuccessful due to low expression of recombinant protein in E.coli. In this project, I am focusing on enhancing the bacterial induction in order to establish robust purification of recombinant Dbp5. This will help in developing ATPase assays involving Dbp5, Nup42, and Gle1. These ATPase assays will aid in better understanding the effects of Nup42 and Gle1 on Dbp5’s ATPase activity and will allow for future study on Dbp5’s ATPase activity. In order to enhance the bacterial induction, E. coli cells were transformed with a GST-Dbp5 plasmid and were induced with varying amounts of IPTG to determine the best procedure for bacterial induction. Results from the bacterial induction have indicated that alternative methods for bacterial induction should be explored. Future experiments will look into further enhancing the bacterial induction of Dbp5 in order to establish a biochemical system analyzing the ATPase activity of GST-Dbp5.
Recommended Citation
Utley, Sarah R.; Rigsby, Rachel E. PhD; and Adams, Rebecca L. PhD, "Establishing a Biochemical System for the Purification and ATPase activity of GST-Dbp5" (2022). Science University Research Symposium (SURS). 13.
https://repository.belmont.edu/surs/13
