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Control of Clminus Efflux in Chara corallina by Cytosolic pH, Free Ca2+, and Phosphorylation Indicates a Role of Plasma Membrane Anion Channels in Cytosolic pH Regulation1

Eva Johannes2, *, Alan Crofts, and Dale Sanders

The Plant Laboratory, Biology Department, University of York, P.O. Box 373, York YO1 5YW, United Kingdom

Enhanced Cl- efflux during acidosis in plants is thought to play a role in cytosolic pH (pHc) homeostasis by short-circuiting the current produced by the electrogenic H+ pump, thereby facilitating enhanced H+ efflux from the cytosol. Using an intracellular perfusion technique, which enables experimental control of medium composition at the cytosolic surface of the plasma membrane of charophyte algae (Chara corallina), we show that lowered pHc activates Cl- efflux via two mechanisms. The first is a direct effect of pHc on Cl- efflux; the second mechanism comprises a pHc-induced increase in affinity for cytosolic free Ca2+ ([Ca2+]c), which also activates Cl- efflux. Cl- efflux was controlled by phosphorylation/dephosphorylation events, which override the responses to both pHc and [Ca2+]c. Whereas phosphorylation (perfusion with the catalytic subunit of protein kinase A in the presence of ATP) resulted in a complete inhibition of Cl- efflux, dephosphorylation (perfusion with alkaline phosphatase) arrested Cl- efflux at 60% of the maximal level in a manner that was both pHc and [Ca2+]c independent. These findings imply that plasma membrane anion channels play a central role in pHc regulation in plants, in addition to their established roles in turgor/volume regulation and signal transduction.

1   This work was supported by the Biotechnology and Biological Sciences Research Council (studentship to A.C. and research fellowship to E.J.).
2   Present address: Department of Botany, Box 7612, North Carolina State University, Raleigh, NC 27695-7612.
*   Corresponding author; e-mail eva_johannes{at}ncsu.edu; fax 1-919-515-3436.

Plant Physiol. (1998) 118: 173-181
Copyright Clearance Center:   0032-0889/98/118//09
© 1998 American Society of Plant Physiologists




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