H-independent glutamine transport in plant root tips.
BACKGROUND
Glutamine is one of the primary amino acids in nitrogen assimilation and often the most abundant amino acid in plant roots. To monitor this important metabolite, a novel genetically encoded fluorescent FRET-reporter was constructed and expressed in Arabidopsis thaliana. As a candidate for the glutamine fluxes, the root tip localized, putative amino acid transporter CAT8 was analyzed and heterologously expressed in yeast and oocytes.


RESULTS
Rapid and reversible in vivo fluorescence changes were observed in reporter-expressing root tips upon exposure and removal of glutamine. FRET changes were detected at acid and neutral pH and in the presence of a protonophore, suggesting that part of the glutamine fluxes were independent of the pH. The putative amino acid transporter CAT8 transported glutamine, had a half maximal activity at approximately 100 microM and the transport was independent of external pH. CAT8 localized not only to the plasma membrane, but additionally to the tonoplast, when tagged with GFP. Ultrastructural analysis confirmed this dual localization and additionally identified CAT8 in membranes of autophagosomes. Loss-of function of CAT8 did not affect growth in various conditions, but over-expressor plants had increased sensitivity to a structural substrate analog, the glutamine synthetase inhibitor L-methionine sulfoximine.


CONCLUSIONS
The combined data suggest that proton-independent glutamine facilitators exist in root tips.
Yang
Huaiyu
H
Plant Biomass and Nutrition, Institute of Botany, Darmstadt University of Technology, Darmstadt, Germany.
Bogner
Martin
M
Stierhof
York-Dieter
YD
Ludewig
Uwe
U
D001692:Biological Transport; D031541:Fluorescence Resonance Energy Transfer; D005973:Glutamine / Q000378:metabolism*; D006859:Hydrogen / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D018517:Plant Roots / Q000378:metabolism*
D005973:Glutamine; D006859:Hydrogen