Distinct transport mechanisms in yeast ammonium transport/sensor proteins of the Mep/Amt/Rh family and impact on filamentation.
Ammonium transport proteins of the Mep/Amt/Rh family include microbial and plant Mep/Amt members, crucial for ammonium scavenging, and animal Rhesus factors likely involved in ammonium disposal. Recent structural information on two bacterial Mep/Amt proteins has revealed the presence, in the hydrophobic conducting pore, of a pair of preserved histidines proposed to play an important role in substrate conductance, by participating either in NH(4)(+) deprotonation or in shaping the pore. Here we highlight the existence of two functional Mep/Amt subfamilies distinguishable according to whether the first of these histidines is conserved, as in yeast ScMep2, or replaced by glutamate, as in ScMep1. Replacement of the native histidine of ScMep2 with glutamate leads to conversion from ScMep2 to ScMep1-like properties. This includes a two-unit upshift of the optimal pH for transport and an increase of the transport rate, consistent with alleviation of an energy-limiting step. Similar effects are observed when the same substitution is introduced into the Escherichia coli AmtB protein. In contrast to ScMep1, ScMep2 is proposed to play an additional signaling role in the induction of filamentous growth, a dimorphic change often associated with virulence in pathogenic fungi. We show here that the histidine to glutamate substitution in ScMep2 leads to uncoupling of the transport and sensor functions, suggesting that a ScMep2-specific transport mechanism might be responsible for filamentation. Our overall data suggest the existence of two functional groups of Mep/Amt-type proteins with different transport mechanisms and distinct impacts on cell physiology and signaling.
Laboratoire de Physiologie Moléculaire de la Cellule, IBMM, the Université Libre de Bruxelles, Rue des Professeurs Jeener et Brachet 12, Gosselies, Belgium.
Mélanie
M
Boeckstaens
Bruno
B
André
Anna Maria
AM
Marini
D000595:Amino Acid Sequence; D001692:Biological Transport; D004926:Escherichia coli / Q000235:genetics; D018698:Glutamic Acid / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D009584:Nitrogen / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction; D013347:Subcellular Fractions / Q000378:metabolism
D000644:Quaternary Ammonium Compounds; D018698:Glutamic Acid; D006639:Histidine; D009584:Nitrogen