DETERMINING THE ROLE OF K63 UBIQUITINATION IN ER TRANSLATION DURING OXIDATIVE STRESS

Oxidative stress is a pervasive environmental challenge linked to the development and progression of several diseases, including cardiovascular disease, cancer, and immunodeficiency. Understanding various cellular responses to oxidative stress is essential in developing therapeutic interventions. The Ubiquitin Proteasome System (UPS) regulates several cellular processes, including protein degradation, synthesis, and gene expression. The Silva lab investigates protein translation by K63 ubiquitination during oxidative stress. Recent literature indicates significant translational regulation occurs in the endoplasmic reticulum (ER), and our lab highlighted preferential K63 ubiquitination of ER-bound ribosomes under oxidative stress in mammalian cells. Despite this, numerous functions of ER-localized translation remain unsolved. This study will determine how stress-induced K63 ubiquitination plays a role in ER-localized translation regulation using saccharomyces cerevisiae (yeast) as a model. To investigate this process, we will utilize several techniques, including cellular fractionation using differential centrifugation. We will further validate differences in accumulation between cellular compartments by performing western blot analysis targeting ubiquitin conjugates. These studies will allow us to better understand whether ER-localized K63 ubiquitination is conserved across species and provide avenues for understanding its essentiality for cellular responses to oxidative stress. Eventually, this will enable us to develop targeted therapies that leverage the power of this cellular modification to better control and alleviate the detrimental effects of oxidative stress and pave the way for future treatments and interventions.