RNA editing in plants converts cytidines to uridines (C-to-U) in chloroplast and
mitochondrial transcripts. Pentatricopeptide repeat (PPR) protein family members have
been shown to be required for editing, and possess key characteristics of editing
deaminases found in other organisms. PPRs have an N-terminal RNA binding domain
responsible for RNA recognition and a C-terminal DYW domain that has deaminase-like
characteristics.
The role of several features of the DYW domain was investigated. The DYW domain
includes the HXE motif that provides a glutamate residue that is catalytically required.
Glutamate to alanine substitution ablated editing, and establishes a key characteristic of
the DYW domain that is required for editing. In addition, a highly conserved PG box was
identified between the N- and C-terminal domains that was required for editing in PPRs
that lacked a DYW domain, and the PG box may be required for protein-protein
interactions to recruit a deaminase in trans. These observations led to the development
of the cis and trans-editing models for RNA editing that posits that the deaminase may be
provided in cis from a single PPR or in trans in PPRs that lack a DYW domain. PPRs, such as MEF8, which have a short RNA binding domain and an intact deaminase domain,were identified as candidates for a trans-deaminases. RNAseq analysis was performed
on mef8 null mutants and catalytically ablated variants to examine the role of MEF8 in
mitochondrial editing. Sixty editing sites were affected, and suggests that MEF8 may
participate as a trans-editing deaminase by providing deaminase capability for a large
number of editing sites.
The role of reactive oxygen species (ROS) during editing dysfunction was investigated to
examine the potential mechanisms of sensing and signaling oxidative distress. LPA66
mutant plants fail to edit a photosystem II polypeptide and exhibit a strong phenotype.
Mutant plants produced elevated levels of ROS, and transcriptomic analysis revealed
higher expression of ROS reactive network genes and PPR or editing related genes.
Lipidomics analysis of the mutant indicated highly elevated levels of oxylipins including
arabidoside A and G and phytoprostanes. Possible signaling mechanisms through
oxylipins and jasmonic acid pathways are discussed.