Genetic dissection of methylcrotonyl CoA carboxylase indicates a complex role for mitochondrial leucine catabolism during seed development and germination.
3-methylcrotonyl CoA carboxylase (MCCase) is a nuclear-encoded, mitochondrial-localized biotin-containing enzyme. The reaction catalyzed by this enzyme is required for leucine (Leu) catabolism, and it may also play a role in the catabolism of isoprenoids and the mevalonate shunt. In Arabidopsis, two MCCase subunits (the biotinylated MCCA subunit and the non-biotinylated MCCB subunit) are each encoded by single genes (At1g03090 and At4g34030, respectively). A reverse genetic approach was used to assess the physiological role of MCCase in plants. We recovered and characterized T-DNA and transposon-tagged knockout alleles of the MCCA and MCCB genes. Metabolite profiling studies indicate that mutations in either MCCA or MCCB block mitochondrial Leu catabolism, as inferred from the increased accumulation of Leu. Under light deprivation conditions, the hyper-accumulation of Leu, 3-methylcrotonyl CoA and isovaleryl CoA indicates that mitochondrial and peroxisomal Leu catabolism pathways are independently regulated. This biochemical block in mitochondrial Leu catabolism is associated with an impaired reproductive growth phenotype, which includes aberrant flower and silique development and decreased seed germination. The decreased seed germination phenotype is only observed for homozygous mutant seeds collected from a parent plant that is itself homozygous, but not from a parent plant that is heterozygous. These characterizations may shed light on the role of catabolic processes in growth and development, an area of plant biology that is poorly understood.
Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011, USA. dimmas@iastate.edu
Geng
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Ding
Ping
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Che
Hilal
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Ilarslan
Eve S
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Wurtele
Basil J
BJ
Nikolau
D000214:Acyl Coenzyme A / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D019735:Carbon-Carbon Ligases / Q000235:genetics* / Q000378:metabolism; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D018525:Germination / Q000235:genetics*; D006720:Homozygote; D007930:Leucine / Q000378:metabolism*; D008855:Microscopy, Electron, Scanning; D008928:Mitochondria / Q000378:metabolism; D016254:Mutagenesis, Insertional; D030821:Plants, Genetically Modified; D021122:Protein Subunits / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
D000214:Acyl Coenzyme A; D029681:Arabidopsis Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D021122:Protein Subunits; C036483:T-DNA; C017447:isovaleryl-coenzyme A; C033618:tiglyl-coenzyme A; D019735:Carbon-Carbon Ligases; C021573:methylcrotonoyl-CoA carboxylase; D007930:Leucine