These results suggest that non pT H ATPase was lost in the evolutionary transition from bryophytes to vascular plants. Regulatory Mechanism of the pT H ATPase in M. polymorpha In vascular plants, phosphorylation of the penultimate Thr of the plasma membrane H ATPase and subsequent binding of the 14 3 3 protein to the phosphorylated C terminus is the most common activation mechanism for the H ATPase . We found that the pT H ATPase in thalli of M. polymorpha is phosphorylated in its penultimate Thr and binds to the 14 3 3 protein in response to FC . These results clearly indicate that the pT H ATPase in M. polymorpha might be activated via an identical mechanism to that in vascular plants. Moreover, we showed that the phosphorylation status of the penultimate Thr of the pT H ATPase in thalli is regulated by phosphorylation in response to physiological signals such as light, Suc, and osmotic shock . Similarly, Suc was reported to induce phosphorylation of the plasma membrane H ATPase in Arabidopsis seedlings , and osmotic shock likely induced phosphorylation of the plasma membrane H ATPase in tomato culture cells , suggesting that the phosphorylation status of the pT H ATPase in the liverwort M.
polymorpha is also regulated by similar physiological signals to those in vascular plants. It should be noted that we measured ATP hydrolytic activity of the H ATPase according to a previous method for vascular plants , but we could not detect increased ATP hydrolytic activity of the H ATPase in response to physiological signals in cell extracts and microsomes from thalli because of high background noise from nonspecific ATP hydrolytic activity in these samples . Further peptide synthesis investigations will be needed to establish measurement methods for plasma membrane H ATPase activity in the liverwort M. polymorpha and to demonstrate that phosphorylation of the penultimate Thr is correlated with the activation status of the H ATPase. In addition, our results suggest that M. polymorpha possesses the identical similar protein kinase and protein phosphatase that directly regulate the phosphorylation status of the pT H ATPase and might have obtained these components in parallel with the evolution of the pT H ATPase.
We should note, however, that the protein kinase and phosphatase have not yet been identified in vascular plants, although they have been extensively investigated . Identification of protein kinase and phosphatase, including Camptothecin those of M. polymorpha, will provide novel understanding for the regulation of pT H ATPase in plants. Photosynthetic Control of the Phosphorylation Status of the pT H ATPase in M. polymorpha In this study, we found that light induces phosphorylation of the H ATPase in thalli and that the photosynthesis inhibitors DCMU and DBMIB inhibit light induced phosphorylation . These results indicate that photosynthesis controls the phosphorylation status of the pT H ATPase in thalli.