Complex assembly and metabolic profiling of Arabidopsis thaliana plants overexpressing vitamin B₆ biosynthesis proteins.
In plants, vitamin B₆ biosynthesis requires the activity of PDX1 and PDX2 proteins. Arabidopsis thaliana encodes for three PDX1 proteins, named PDX1.1, 1.2, and 1.3, but only one PDX2. Here, we show in planta complex assembly of PDX proteins, based on split-YFP and FPLC assays, and can demonstrate their presence in higher complexes of around 750 kDa. Metabolic profiling of plants ectopically expressing the different PDX proteins indicates a negative influence of PDX1.2 on vitamin B₆ biosynthesis and a correlation between aberrant vitamin B6 content, PDX1 gene expression, and light sensitivity specifically for PDX1.3. These findings provide first insights into in planta vitamin B₆ synthase complex assembly and new information on how the different PDX proteins affect plant metabolism.
Leuendorf
Jan Erik
JE
Angewandte Genetik, Freie Universität Berlin, 14195 Berlin, Germany.
Osorio
Sonia
S
Szewczyk
Agnieszka
A
Fernie
Alisdair R
AR
Hellmann
Hanjo
H
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D019759:Carbon-Nitrogen Lyases; D002850:Chromatography, Gel; D019884:Nitrogenous Group Transferases / Q000235:genetics / Q000378:metabolism; D025101:Vitamin B 6 / Q000096:biosynthesis* / Q000235:genetics
D029681:Arabidopsis Proteins; C000588558:PDX1.2 protein, Arabidopsis; D025101:Vitamin B 6; D019884:Nitrogenous Group Transferases; D019759:Carbon-Nitrogen Lyases; C505161:At2g38230 protein, Arabidopsis