Plant J. 2021 Nov 07.
In the chloroplast, OZ1 is a RanBP2-type zinc finger protein required for many RNA editing events, a process by which specific cytosines are enzymatically converted to uracils as a correction mechanism for missense mutations in the organelle genomes. RNA editing is carried out by a large multi-protein complex called the "editosome" that contains members of the PPR protein family, the RIP/MORF family, and the ORRM family in addition to OZ1. OZ1 is an 82-kDa protein with distinct domains, including a pair of zinc finger domains and a unique C-terminal region. To elucidate the functions of these domains, we have generated truncations of OZ1 for use in protein-protein interaction assays that identified the C-terminal region of OZ1 as well as the zinc finger domains as the primary interactors with PPR proteins, which are factors required for site-specificity and enzymatic editing. Expression of these OZ1 truncations in vivo showed that the zinc finger domains were required to restore chloroplast RNA editing in oz1 knockout plants. Mutation of key structural residues in the zinc finger domains showed that they are necessary for editing and required for interaction with ORRM1, a general editing factor with an RNA-binding domain. These functional characterizations of the zinc fingers and novel C-terminal domain contribute to our understanding of the model for the chloroplast plant editosome.
Keywords: RNA editing; chloroplast; plant editosome; zinc finger