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J Biol Chem, Vol. 273, Issue 47, 31463-31470, November 20, 1998
From the Instituto de Bioquímica Vegetal y
Fotosíntesis, Consejo Superior de Investigaciones
Científicas-Universidad de Sevilla, E-41092
Sevilla, Spain
Ammonium is an important nitrogen source for many
microorganisms and plants. Ammonium transporters whose activity can be
probed with [14C]methylammonium have been described
in several organisms including some cyanobacteria, and amt
genes encoding ammonium/methylammonium permeases have been recently
identified in yeast, Arabidopsis thaliana, and some
bacteria. The unicellular cyanobacterium Synechocystis sp.
PCC 6803 exhibited a [14C]methylammonium uptake activity
that was inhibited by externally added ammonium. Three putative
amt genes that are found in the recently published complete
sequence of the chromosome of strain PCC 6803 were inactivated by
insertion of antibiotic resistance-encoding gene-cassettes. The
corresponding mutant strains were impaired in uptake of
[14C]methylammonium. Open reading frame
sll0108 (amt1) was responsible for a high
affinity uptake activity (Ks for methylammonium, 2.7 µM), whereas open reading frames sll1017
(amt2) and sll0537 (amt3) made
minor contributions to uptake at low substrate concentrations. Expression of the three amt genes was higher in
nitrogen-starved cells than in cells incubated in the presence of a
source of nitrogen (either ammonium or nitrate), but amt1
was expressed at higher levels than the other two amt
genes. Transcription of amt1 was found to take place from a
promoter bearing the structure of the cyanobacterial promoters
activated by the nitrogen control transcription factor, NtcA.
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