Date of Graduation

5-2025

Document Type

Dissertation

Degree Name

Doctor of Philosophy in Cell & Molecular Biology (PhD)

Degree Level

Graduate

Department

Cell & Molecular Biology

Advisor/Mentor

Lewis, Jeffrey A.

Committee Member

Lessner, Daniel J.

Second Committee Member

Du, Yuchun

Third Committee Member

Fan, Chenguang

Keywords

hog1; mkt1; proteomics; stress

Abstract

The stress response of Saccharomyces cerevisiae is a valuable area of study for understanding cellular adaptation mechanisms that govern survival in fluctuating environments. This dissertation investigates stress response regulation in yeast through the roles of Hog1 mitogen-activated protein kinase (MAPK) and the post-transcriptional regulator Mkt1, a key post-transcriptional regulator, in diverse wild yeast isolates. Hog1 is traditionally known for its central role in osmotic stress regulation in the high-osmolarity glycerol (HOG) pathway. This study expands the functional scope of Hog1 to include oxidative and ethanol-induced stress responses.

In parallel, we investigate post-transcriptional regulation of stress responses by the Mkt1 interactome, linking Mkt1 to metabolic and translational regulation. Using SILAC-based proteomics, we outline the dynamic nature of the Mkt1 interactome.

We further address the pitfalls of ribonucleoprotein complex procedures in RNA-protein interaction studies, proposing enhanced protocols for more comprehensive investigations into stress-induced gene regulation.

Collectively, this dissertation offers new knowledge on the complexity of stress adaptation in S. cerevisiae, and the importance of strain-specific variation within stress response pathways, demonstrating the value of studying natural variation in molecular studies. The work not only advances the understanding of Hog1 and Mkt1 in stress regulation but also gives an overall perspective of how natural genetic variation influences cellular responses to environmental stress.

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