Systematic Phenotypic Screen of Arabidopsis Peroxisomal Mutants Identifies Proteins Involved in β-Oxidation

Peroxisomes are highly dynamic and multifunctional organelles essential to development. Plant peroxisomes accommodate a multitude of metabolic reactions, many of which are related to the β-oxidation of fatty acids or fatty acid-related metabolites. Recently, several dozens of novel peroxisomal prote...

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Bibliographic Details
Published in:Plant physiology (Bethesda) 2014-11, Vol.166 (3), p.1546-1559
Main Authors: Cassin-Ross, Gaëlle, Hu, Jianping
Format: Article
Language:English
Subjects:
2,4-Dichlorophenoxyacetic Acid - analogs & derivatives
2,4-Dichlorophenoxyacetic Acid - metabolism
2,4-Dichlorophenoxyacetic Acid - pharmacology
Arabidopsis - drug effects
Arabidopsis - genetics
Arabidopsis - growth & development
Arabidopsis - metabolism
Arabidopsis Proteins - genetics
Arabidopsis Proteins - metabolism
BASIC BIOLOGICAL SCIENCES
Carbohydrate Metabolism - genetics
Enzymes
Fatty acids
Fatty Acids - metabolism
Fatty Acids, Unsaturated - metabolism
Gene Expression Regulation, Plant
Germination
Germination - genetics
Isocitrate Dehydrogenase - genetics
Isocitrate Dehydrogenase - metabolism
Lipid bodies
Lipid Droplets - metabolism
Membrane Proteins - genetics
Membrane Proteins - metabolism
Mutation
Oxidation-Reduction
Peroxisomes
Peroxisomes - genetics
Peroxisomes - metabolism
Phenotype
Phenotypes
Plants
Repressor Proteins - genetics
Repressor Proteins - metabolism
Seed germination
Seedlings
Seeds - growth & development
Seeds - metabolism
Statistical significance
SYSTEMS AND SYNTHETIC BIOLOGY
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Summary:Peroxisomes are highly dynamic and multifunctional organelles essential to development. Plant peroxisomes accommodate a multitude of metabolic reactions, many of which are related to the β-oxidation of fatty acids or fatty acid-related metabolites. Recently, several dozens of novel peroxisomal proteins have been identified from Arabidopsis (Arabidopsis thaliana) through in silico and experimental proteomic analyses followed by in vivo protein targeting validations. To determine the functions of these proteins, we interrogated their transfer DNA insertion mutants with a series of physiological, cytological, and biochemical assays to reveal peroxisomal deficiencies. Sugar dependence and 2,4-dichlorophenoxybutyric acid and 12-oxo-phytodienoic acid response assays uncovered statistically significant phenotypes in β-oxidation-related processes in mutants for 20 of 27 genes tested. Additional investigations uncovered a subset of these mutants with abnormal seed germination, accumulation of oil bodies, and delayed degradation of long-chain fatty acids during early seedling development. Mutants for seven genes exhibited deficiencies in multiple assays, strongly suggesting the involvement of their gene products in peroxisomal β-oxidation and initial seedling growth. Proteins identified included isoforms of enzymes related to β-oxidation, such as acyl-CoA thioesterase2, acyl-activating enzyme isoforml, and acyl-activating enzyme isoform5, and proteins with functions previously unknown to be associated with β-oxidation, such as Indigoidine synthase A, Senescence-associated protein/B12D-related protein1, Betaine aldehyde dehydrogenase, and Unknown protein5. This multipronged phenotypic screen allowed us to reveal β-oxidation proteins that have not been discovered by single assay-based mutant screens and enabled the functional dissection of different isoforms of multigene families involved in β-oxidation.
ISSN:0032-0889
1532-2548
1532-2548
DOI:10.1104/pp.114.250183