Date of Award
Summer 2025
Document Type
Open Access Thesis
Department
Chemistry and Biochemistry
First Advisor
F. Wayne Outten
Abstract
Iron-sulfur (Fe-S) clusters are an essential cofactor for a wide range of cellular processes, and their assembly and storage require tight regulation.
In Escherichia coli (E. coli), there are two pathways of Fe-S cluster assembly, the Isc and the Suf pathways. The Isc pathway is used under good growth conditions, and the Suf pathway is used under iron starvation and oxidative stress conditions. Once the cluster has been assembled on the scaffold protein of either pathway, it is transferred to Fe-S cluster trafficking proteins. In E. coli, the monothiol glutaredoxin GrxD and the BolA-type proteins BolA and IbaG have been implicated in Fe-S cluster trafficking and iron homeostasis. GrxD can form heterodimers with either of the BolA-type proteins. GrxD has also been shown to interact with the scaffold protein of the Isc pathway, IscU, and the cluster trafficking protein ErpA, and published results from our lab have indicated a cluster storage role for GrxD. This research utilizes proximity labeling, qPCR, and cell growth assays to further define the role of GrxD, specify the purpose of the GrxD heterodimers with the BolA-type proteins, and learn more about the Suf pathway of Fe-S cluster biosynthesis by studying the effect of point mutations on Fe-S cluster assembly.
The results show that the GrxD-IbaG heterodimer is important for maintaining the labile iron pool of the cell, and amino acids K9 and R16 of SufE are essential for Fe-S cluster formation via the Suf pathway.
Rights
© 2025, Dexter McLaurin Reasons
Recommended Citation
Reasons, D. M.(2025). In Vivo Analysis of FE-S Cluster Assembly and Iron Homeostasis in Escherichia Coli. (Master's thesis). Retrieved from https://scholarcommons.sc.edu/etd/8534