Induction of vacuolar ATPase and mitochondrial ATP synthase by aluminum in an aluminum-resistant cultivar of wheat

Two 51-kD aluminum (Al)-induced proteins (RMP51, root membrane proteins of 51 kD) were recently discovered in an aluminum-resistant cultivar of wheat (Triticum aestivum) cv PT741 (Basu et al., 1994a). These proteins segregate with the aluminum resistance phenotype in a segregating population arising from a cross between Al-resistant cv PT741 and Al-sensitive cv Katepwa (Taylor et al., 1997). The proteins have been purified by continuous elution electrophoresis and analyzed by peptide microsequencing. Sequence analysis of the purified peptides revealed that they are homologous to the B subunit of the vacuolar H+-ATPase (V-ATPase) and the alpha- and beta -subunits of the mitochondrial ATP synthase (F1F0-ATPase). To confirm that these ATPases are induced by Al, ATPase activity and transcript levels were analyzed under Al stress. Both V-ATPase and F1F0-ATPase activities were induced by Al and responded in a dose-dependent manner to 0 to 150 muM Al. In contrast, plasma membrane H+-ATPase (P-ATPase) activity decreased to 0.5x control levels, even when plants were exposed to 25 muM Al. Northern analysis showed that the transcript encoding the B subunit of V-ATPase increased by 2.2x in a dose-dependent manner, whereas levels of the transcript encoding the alpha -subunit of F1F0-ATPase remained constant. The effect of Al on ATPase activity in other cultivars was also examined. The Al-resistant cultivar, cv PT741, was the only cultivar to show induction of V- and F1F0-ATPases. These results suggest that the V-ATPase in cv PT741 is responding specifically to Al stress with the ATP required for its activity supplied by ATP synthase to maintain energy balance within the cell.