title: Identification of two pentatricopeptide repeat genes required for RNA editing and zinc binding by C-terminal cytidine deaminase-like domains.
abstract: Many transcripts expressed from plant organelle genomes are modified by C-to-U RNA editing. Nuclear encoded pentatricopeptide repeat (PPR) proteins are required as RNA binding specificity determinants in the RNA editing mechanism. Bioinformatic analysis has shown that most of the Arabidopsis PPR proteins necessary for RNA editing events include a C-terminal portion that shares structural characteristics with a superfamily of deaminases. The DYW deaminase domain includes a highly conserved zinc binding motif that shares characteristics with cytidine deaminases. The Arabidopsis PPR genes, ELI1 and DOT4, both have DYW deaminase domains and are required for single RNA editing events in chloroplasts. The ELI1 DYW deaminase domain was expressed as a recombinant protein in Escherichia coli and was shown to bind two zinc atoms per polypeptide. Thus, the DYW deaminase domain binds a zinc metal ion, as expected for a cytidine deaminase, and is potentially the catalytic component of an editing complex. Genetic complementation experiments demonstrate that large portions of the DYW deaminase domain of ELI1 may be eliminated, but the truncated genes retain the ability to restore editing site conversion in a mutant plant. These results suggest that the catalytic activity can be supplied in trans by uncharacterized protein(s) of the editosome.
authors.affiliation: From the Departments of Developmental and Cell Biology and.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Hayes
authors.forename: Karolyn
authors.initials: K
authors.lastname: Giang
authors.forename: Beniam
authors.initials: B
authors.lastname: Berhane
authors.forename: R Michael
authors.initials: RM
authors.lastname: Mulligan
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D001937:Brassica / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003564:Cytidine Deaminase / Q000737:chemistry*; D017343:Genes, Plant; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017393:RNA Editing; D015032:Zinc / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; C538752:pentatricopeptide repeat protein, Arabidopsis; D003564:Cytidine Deaminase; D015032:Zinc
keywords: Chloroplast; Gene Expression; Mass Spectrometry (MS); Metalloenzymes; RNA Modification