Date of Award

Spring 2022

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Molecular Biophysics and Biochemistry

First Advisor

De La Cruz, Enrique

Abstract

Dbp5 from Saccharomyces cerevisiae is an essential DEAD-box protein required for many aspects of mRNA metabolism including nuclear mRNA export. The ATPase cycle of Dbp5 is required for its in vivo function in mRNA export. The Dbp5 ATPase cycling kinetics dictate how the energy from ATP binding and hydrolysis is coupled to RNA remodeling. Several factors are known to regulate Dbp5 ATPase and mRNA export activity including the small molecule inositol hexaphosphate (InsP6), RNA, and the nucleoporins Gle1 and Nup159. In order to quantify how these regulatory factors, influence the ATPase activity and functionality of Dbp5, we measured the rate and equilibrium constants of the Dbp5 ATPase alone and in conjunction with these regulatory factors using various steady-state and transient kinetic techniques. Knowledge of how Nup159, Gle1, and RNA modulate the Dbp5 ATPase kinetics allows us to identify the mechanism of Dbp5 mediated mRNA export and regulation.

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