yeast in these conditions. The Alr1p and Alr2p proteins consist of 859 and 858 amino acids respectively and are 70% identical. In a region in the C-terminal, half of these proteins are weakly similar to the full CorA protein. The computer-predicted TM topology of Alr1p is shown in the figure. The presence of a third TM domain was suggested by MacDiarmid and Gardner (1998), on the strength on sequence homology, and more recently by Lee and Gardner (2006), on the strength of mutagenesis studies, making the TM topology of these proteins more like that of CorA (see figure). Also, Alr1p contains the conserved GMN motif at the outside end of TM 2 (TM 2') and the mutation of the methionine (M) in this motif to a leucine (L) led to the loss of transport capability. The figure shows the two possible TM topologies of Alr1p. Part A of the figure shows the computer-predicted membrane topology of the Alr1p protein in yeast and part B shows the topology of Alr1p based on the experimental results of Lee and Gardner (2006). The GMN motif location is indicated in red and the TM domains in light blue. The orientation in the membrane and the positions of the N- and C-termini are indicated, the various sizes of the soluble domains are given in amino acids (AA), and TM domains are numbered by their similarity to CorA. Where any TM domain is missing, the remaining domains are numbered with primes. The figure is not drawn to scale. A third ALR-like gene is present in S. cerevisiae and there are two homologous genes in both Schizosaccharomyces pombe and Neurospora crassa. These proteins contain a GMN motif like that of CorA, with the exception of the second N. crassa gene. No ALR-like genes have been identified in species outside of the fungi. Membrane fractionation and green fluorescent protein (GFP) fusion studies established that Alr1p is localised to the plasma membrane. The localisation of the Alr1p was observed to be internalised and degraded in the vacuole in response to extracellular cations. Mg2+, at very low extracellular concentrations (100 μM; < 10% of the standard media Mg2+ content), and Co2+ and Mn2+ at relatively high concentrations (> 20× standard media), induced the change in Alr1p protein localisation, and the effect was dependent on functional ubiquitination, endocytosis and vacuolar degradation. This mechanism was proposed to allow the regulation of Mg2+