title: Analysis of Arabidopsis Accessions Hypersensitive to a Loss of Chloroplast Translation.
abstract: Natural accessions of Arabidopsis (Arabidopsis thaliana) differ in their ability to tolerate a loss of chloroplast translation. These differences can be attributed in part to variation in a duplicated nuclear gene (ACC2) that targets homomeric acetyl-coenzyme A carboxylase (ACCase) to plastids. This functional redundancy allows limited fatty acid biosynthesis to occur in the absence of heteromeric ACCase, which is encoded in part by the plastid genome. In the presence of functional ACC2, tolerant alleles of several nuclear genes, not yet identified, enhance the growth of seedlings and embryos disrupted in chloroplast translation. ACC2 knockout mutants, by contrast, are hypersensitive. Here we describe an expanded search for hypersensitive accessions of Arabidopsis, evaluate whether all of these accessions are defective in ACC2, and characterize genotype-to-phenotype relationships for homomeric ACCase variants identified among 855 accessions with sequenced genomes. Null alleles with ACC2 nonsense mutations, frameshift mutations, small deletions, genomic rearrangements, and defects in RNA splicing are included among the most sensitive accessions examined. By contrast, most missense mutations affecting highly conserved residues failed to eliminate ACC2 function. Several accessions were identified where sensitivity could not be attributed to a defect in either ACC2 or Tic20-IV, the chloroplast membrane channel required for ACC2 uptake. Overall, these results underscore the central role of ACC2 in mediating Arabidopsis response to a loss of chloroplast translation, highlight future applications of this system to analyzing chloroplast protein import, and provide valuable insights into the mutational landscape of an important metabolic enzyme that is highly conserved throughout eukaryotes.
authors.affiliation: Department of Plant Biology, Ecology, and Evolution, Oklahoma State University, Stillwater, Oklahoma 74078.
authors.forename: Nicole
authors.initials: N
authors.lastname: Parker
authors.affiliation: Department of Plant Biology, Ecology, and Evolution, Oklahoma State University, Stillwater, Oklahoma 74078.
authors.forename: Yixing
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Plant Biology, Ecology, and Evolution, Oklahoma State University, Stillwater, Oklahoma 74078 meinke@okstate.edu.
authors.forename: David
authors.initials: D
authors.lastname: Meinke
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics; D001483:Base Sequence; D002736:Chloroplasts / Q000378:metabolism*; D017124:Conserved Sequence; D060146:Ecotype; D055786:Gene Knockout Techniques; D005820:Genetic Testing; D018525:Germination / Q000187:drug effects / Q000235:genetics; D020125:Mutation, Missense / Q000235:genetics; D014176:Protein Biosynthesis; D059386:Reverse Genetics; D036226:Seedlings / Q000187:drug effects / Q000235:genetics; D000198:Spectinomycin / Q000494:pharmacology
chemical_list: D029681:Arabidopsis Proteins; D000198:Spectinomycin