Characterization of ribosome-associated mRNA species degraded by autophagy. Saccharomyces cerevisiae BY4741 strain:X2180-1B

Background: Autophagy is a bulk degradation system of cellular components induced by nutrient starvation. The role of autophagy in the degradation of proteins and organelles is well-characterized. However, autophagy-mediated RNA degradation in response to stress and the potential preference of specific RNAs to undergo autophagy-mediated degradation have not been examined. Recently, we revealed that autophagy preferentially delivers a subset of mRNA species to the vacuole. We also found that the selectivity of autophagy-mediated mRNA degradation is coupled with ribosomal association and depends on Atg24 (sorting nexin complex) and Rsa1 (ribosome assembly factor). Aim: In this project, we propose to characterize Atg24- and Rsa1-dependent mRNA species degraded by autophagy. Based on the RNA-seq data, we would like to elucidate the molecular mechanisms and physiological roles of autophagy-mediated Atg24- and Rsa1-dependent mRNA degradation. Samples: Total lysate and Isolated vacuoles from Rny1 (vacuolar RNase) KO cells (RNA degradation defective), Rny1 and Atg24 double KO cells (ribosome-associated mRNA degradation defective), and Rny1 and Rsa1 double KO cells.