Understanding the Localization of ADARs

Adenosine-to-inosine (A-to-I) dsRNA editing is a conserved RNA modification occurs in metazoan including in Caenorhabditis elegans (C. elegans). The editing is catalyzed by ADAR enzymes. In C. elegans, there are two ADAR genes, ADR-1 and ADR-2. ADR-1 is a regulator of ADR-2 and ADR-2 is the only active enzyme, which catalyzes the deamination. While lack of editing is lethal for mammals, C. elegans can survive when the editing levels are low, making them a very important model organism for understanding A-to-I editing. ADBP-1 (ADR-2 binding protein-1) is involved in the localization of ADR-2 to the nucleus. We found that in the absence of ADBP-1, the majority of ADR-2 is localized in the cytoplasm. However, ADBP-1 does not have NLS, and how it regulates ADR-2 localization is currently unknown. Our initial results suggest that importins might be involved. Our aim is to understand how ADR-2 localization is determined and regulated, and whether ADBP-1 has other roles apart from regulating ADAR-2 localization. We plan to study where ADR-2 and ADBP-1 bind to each other. We also plan to track both protein localization using tagged CRISPR strains. We will generate adbp-1 deletion strain and characterize the phenotypes. In addition, we will check if importins are involved in the localization of the proteins. Our results will reveal how the localization of these proteins is regulated and shed light on the regulation of A-to-I process and its functions.