Date Approved
2021
Degree Type
Open Access Senior Honors Thesis
Department or School
Chemistry
First Advisor
Steven Backues, Ph.D.
Second Advisor
Hedeel Evans, Ph.D.
Third Advisor
Debbie Heyl-Clegg, Ph.D.
Abstract
Autophagy is a highly conserved eukaryotic cellular recycling process in which the degradation of proteins and cytoplasmic organelles occurs to recycle broken down products throughout the cell. This process plays a critical role in cell survival and maintenance. Once initiated by either nutrient deficiency (bulk autophagy) or recognition of damaged organelles (selective autophagy) this process is mediated by autophagy-related proteins (Atg proteins). The Atg proteins function to form a double-membrane vesicle, the autophagosome, to target part of the cytoplasm and deliver it to the vesicle for breakdown. In bulk autophagy, the presence of these proteins at the phagophore assembly site (PAS) are essential to determining the size and number of autophagosomes formed. There are two main ubiquitin conjugation systems that are known to contribute to the size and number of autophagosomes, Atg12 and Atg8. Atg12 is conjugated to Atg5 by Atg7, an E1-like protein, and Atg10, and E2-like protein. Similarly, Atg8 is conjugated to phosphatidylethanolamine, PE, by Atg 7 and Atg3. How the E2-like protein Atg10 influences the size of autophagosomes formed is not clear. Here we attempt to create hemagglutinin tagged Atg12 and Atg5 in order to determine levels of Atg12-5 conjugation as a readout of Atg10 function. We generated a recombinant yeast strain that expresses detectable levels of Atg5-HA that can be used to determine Atg12-5 conjugation by various Atg10 mutants.
Recommended Citation
Brydie, Ayanda, "How does Atg 10 regulate the autophagic process? An investigation into the Atg12-5 conjugation pathway" (2021). Senior Honors Theses and Projects. 842.
https://commons.emich.edu/honors/842
