A Novel Approach to Establish a Comprehensive Atlas of Human Mitochondrial mRNA Binding Proteins
Abstract
Fiscal Year 2018 Peer Reviewed Medical Research Program Topic Area: Mitochondrial Disease Mitochondria are semi-autonomous organelles that originate from a symbiotic relationship between a protoeukaryotic cell and a primitive oxygen-breathing bacterium. Along evolution, most genes of the primitive prokaryotic endosymbiont were either lost or transferred to the nucleus of the host cell. However, mitochondria still retain their own genome (mtDNA), which encodes for a handful of proteins, 13 in humans, essential components of the oxidative phosphorylation (OXPHOS) system, two ribosomal RNAs (human 12S and 16S rRNAs) and 22 tRNAs, required for mitochondrial translation. The OXPHOS system is responsible for the production of most cellular energy, in the form of ATP molecules. Thus, mtDNA expression is essential for cellular energy production, and its alteration leads to major energetic deficits in cells and tissues, which result in severe human diseases, namely mitochondrial disorders, such as encephalomyopathies and cardiomyopathies. In human mitochondria, control of gene expression appears to occur mainly at the post-transcriptional level through the interplay of mRNAs with RNA-binding proteins (RBPs). Although increasing efforts are being devoted to better understand mitochondrial RBPs (mt-RBPs), the knowledge regarding the roles play by mtRBPs in processes such as initiation of translation or regulation of mRNA stability remains largely incomplete. Moreover, much of our current knowledge of mt-mRNA-protein interactions has been accumulated stepwise over the last two decades, and a systematic study of mt-RBPs is lacking. The biomedical significance of mitochondrial RBPs (mt-RBPs) stems from the fact that defects in mt-mRNA processing, stability or expression, result in an array of human mitochondrial disorders. Insights into these processes and the factors involved are therefore an essential requirement to identify potential targets of therapeutic intervention for the management of mitochondrial diseases. Therefore, here we propose the development of a novel methodology to identify single mRNA-specific RBPs through the combination of mt-mRNA-specific pull-down approaches. The methodology we propose to implement is based on the construction of Pumilio-derived RNA-binding domains targeted to mitochondria and capable of specifically recognizing individual mt-mRNAs. Our long-term goal is to establish an atlas of human mitochondrial mRNA-binding proteins to gain insights into mt-mRNA biology.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Mar 05, 2019
- Source ID
- W81XWH1910124XX0
Entities
People
- Flavia Fontanesi
Organizations
- United States Army
- University of Miami