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Biochemical and Functional Characterization of a Mitochondrial Citrate Carrier in Arabidopsis thaliana
- Source :
- Biochemical Journal
- Publication Year :
- 2020
-
Abstract
- A homolog of the mitochondrial succinate/fumarate carrier from yeast (Sfc1p) has been found in the Arabidopsis genome, named AtSFC1. The AtSFC1 gene was expressed in Escherichia coli, and the gene product was purified and reconstituted in liposomes. Its transport properties and kinetic parameters demonstrated that AtSFC1 transports citrate, isocitrate and aconitate and, to a lesser extent, succinate and fumarate. This carrier catalyzes a fast counter-exchange transport as well as a low uniport of substrates, exhibits a higher transport affinity for tricarboxylates than dicarboxylates, and is inhibited by pyridoxal 5′-phosphate and other inhibitors of mitochondrial carriers to various degrees. Gene expression analysis indicated that the AtSFC1 transcript is mainly present in heterotrophic tissues, and fusion with a green-fluorescent protein localized AtSFC1 to the mitochondria. Furthermore, 35S-AtSFC1 antisense lines were generated and characterized at metabolic and physiological levels in different organs and at various developmental stages. Lower expression of AtSFC1 reduced seed germination and impaired radicle growth, a phenotype that was related to reduced respiration rate. These findings demonstrate that AtSFC1 might be involved in storage oil mobilization at the early stages of seedling growth and in nitrogen assimilation in root tissue by catalyzing citrate/isocitrate or citrate/succinate exchanges.
- Subjects :
- 0106 biological sciences
Nitrogen
Genes, Fungal
Saccharomyces cerevisiae
Arabidopsis
Gene Expression
Mitochondrion
Genes, Plant
medicine.disease_cause
01 natural sciences
Biochemistry
Mitochondrial Proteins
Gene product
03 medical and health sciences
Fumarates
Gene expression
medicine
Arabidopsis thaliana
Molecular Biology
Escherichia coli
030304 developmental biology
Dicarboxylic Acid Transporters
0303 health sciences
biology
Chemistry
Fatty Acids
Tricarboxylic Acids
Biological Transport
Succinates
Cell Biology
biology.organism_classification
Mitochondrial carrier
Mitochondria
Kinetics
Seedlings
Liposomes
Carrier Proteins
010606 plant biology & botany
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Biochemical Journal
- Accession number :
- edsair.doi.dedup.....103dec8da187e4859db0aef92ff4bf64