Predicted topology model and intracellular localization of SpAvt3p.

<p>(A)<i>Top</i>, predicted topology model of SpAvt3p. <i>Bottom</i>, sequence alignments of SpAvt3p (Q10074.1) in TM6 (amino acids 451–477) and analogous regions in the homologs according to CLUSTALW: <i>Saccharomyces cerevisiae</i> Avt3p and Avt4p (P36062 and P50944, respectively), <i>Arabidopsis thaliana</i> At5G65990 (ABH04593), and human hPAT1and hPAT2 (AAI36439 and AAI01104, respectively). Identical and similar residues are denoted by <i>black boxes</i> and <i>gray boxes</i>, respectively. The conserved glutamate residue is indicated by an asterisk. (B) The <i>avt3</i>Δ mutant cells expressing GFP-SpAvt3p fusion protein were subjected to fluorescence microscopy. Vacuolar membranes were stained with FM4-64. BF, bright field; bar, 5 μm.