Expression cloning in yeast of a cDNA encoding a broad specificity amino acid permease from Arabidopsis thaliana

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 90; no. 13; pp. 5944 - 5948
• Main Authors: Frommer, W.B, Hummel, S, Riesmeier, J.W
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01-07-1993; National Acad Sciences; National Academy of Sciences
• Subjects: ADN; Amino Acid Transport Systems; Amino acids; Arabidopsis - enzymology; Arabidopsis - genetics; Arabidopsis - growth & development; ARABIDOPSIS THALIANA; Base Sequence; Biochemistry; Biological and medical sciences; Biological Transport; Botany; Cell physiology; CLONACION; CLONAGE; Cloning, Molecular; CODE GENETIQUE; CODIGO GENETICO; Complementary DNA; Complementation; EXPRESION GENICA; EXPRESSION DES GENES; Flowers & plants; Fundamental and applied biological sciences. Psychology; Genes, Plant; Genetic Complementation Test; Leaves; Membrane and intracellular transports; Membrane transport proteins; Membrane Transport Proteins - genetics; Membrane Transport Proteins - metabolism; Metabolism; Metabolism. Physicochemical requirements; Molecular and cellular biology; Molecular Sequence Data; Nitrogen; Phloem; Plant physiology and development; Plants; Proline - metabolism; SECUENCIA NUCLEICA; Sequence Analysis, DNA; SEQUENCE NUCLEIQUE; Substrate Specificity; Yeasts; Yeasts - enzymology; Yeasts - genetics; Heterologous system; Yeast; Transcription promoter; Gene expression; Fungi; Arabidopsis thaliana; Specificity; Complementary DNA; Cruciferae; Membrane transport; Dicotyledones; Aminoacid; Angiospermae; Ascomycetes; Spermatophyta; Molecular cloning; Saccharomyces cerevisiae; Thallophyta
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.90.13.5944

To study amino acid transport in plants at the molecular level, we have isolated an amino acid permease cDNA from Arabidopsis thaliana by complementation of a yeast mutant defective in proline uptake with a cDNA. The predicted polypeptide of 53 kDa is highly hydrophobic with 12 putative membrane-spanning regions and shows no significant homologies to other known transporters. Expression of the cDNA enables the yeast mutant to take up L-[14C]proline. Competition studies argue for a broad but stereospecific substrate recognition by the permease, which resembles neutral or general amino acid transport systems from Chlorella and higher plants. Both pH dependence and inhibition by protonophores are consistent with a proton symport mechanism.