Am. Marini et al., Cross-talk between ammonium transporters in yeast and interference by the soybean SAT1 protein, MOL MICROB, 35(2), 2000, pp. 378-385
Ammonium uptake in the yeast Saccharomyces cerevisiae involves three membra
ne transporters (Mep1, -2 and -3) belonging to an evolutionarily conserved
protein family that also includes the rhesus (Rh) blood group polypeptides
of erythrocytes. We show here that, in the 26972c mutant defective in NH4transport, the Mep1 protein carrying an amino acid substitution in its cyto
plasmic C-terminus trans-inhibits the closely related Mep3 protein. The sam
e mutation introduced into Mep3 leads to loss of transport activity and thi
s inactive form also trans-inhibits native Mep3. Inhibition of Mep3 is post
-translational and can be overcome by overexpression. These results are con
sistent with a direct interaction between Mep proteins, as is the case for
the Rh polypeptides. The soybean GmSAT1 gene, recently cloned for its abili
ty to complement the NH4+ transport defect of strain 26972c, has been descr
ibed as an NH4+ channel protein involved in the transfer of fixed nitrogen
from the bacteroid to the host plant. We show here that GmSAT1 contains a s
equence homologous to the DNA-binding domain of basic helix-loop-helix (bHL
H) transcription factors. We also show that GmSAT1 restores NH4+ uptake in
the yeast mutant by interfering with the inhibition of Mep3. Our results ar
e not consistent with a direct role of GmSAT1 in ammonium transport.