Abstract

RNA-binding proteins (RBPs) play essential roles in the post-transcriptional control of gene expression. The recent introduction of proteome-wide approaches has dramatically expanded the repertoire of proteins interacting with RNA, revealing many “unconventional” RBPs with other well-established functions, such as metabolic enzymes. Nevertheless, while the repertoire of RBPs is steadily increasing, very little is known about the reconfiguration of the RNA-protein interactions upon stress and RNA-related functions of “unconventional” RBPs. To address this lack of knowledge, we monitored the changes of the mRNA-binding proteome (mRBPome) upon oxidative stress in the yeast Saccharomyces cerevisiae. Intriguingly, we observed prime changes in the RNA associations among enzymes acting in carbon metabolism, which is reminiscent to previously reported metabolic reconfigurations. In our proposed research, we wish to investigate whether the observed changes of enzyme-mRNA interactions are stress-dependent, and undertake a functional analysis of selected enzymes. Therefore, we will i) profile the changes of the mRNA-protein interactome across distinct environmental stress conditions, ii) comprehensively profile the RNA targets and proteins interacting with selected enzymes in stressed and non-stressed cells; and iii) monitor the implications of specific enzyme-RNA interactions in gene expression, for enzyme activity and consequences in stress adaptation. Our research is expected to elucidate “moonlighting” functions of key metabolic enzymes and likely uncovers new principles for cellular stress adaptation. Since both, enzymes acting in carbon metabolism and the cell’s response to oxidative stress are of considerable interest – ranging from improving fermentation processes in yeast which are relevant in the food and biofuel industry to the development of new strategies for cancer treatment – our research could eventually generate economical and societal impact.