AMINO-TERMINUS AND THE FIRST 4 MEMBRANE-SPANNING SEGMENTS OF THE ARABIDOPSIS K+ CHANNEL KAT1 CONFER INWARD-RECTIFICATION PROPERTY OF PLANT-ANIMAL CHIMERIC CHANNELS

The Arabidopsis hyperpolarization-activated (inward-rectifying) K+ channel KAT1 is structurally more similar to animal depolarization-activated (outward-rectifying) K+ channels than to animal hyperpolarization-activated K+ channels. To gain insight into the structural basis for the opposite voltage dependences of plant inward-rectifying and animal outward-rectifying K+ channels, we constructed recombinant chimeric channels between the hyperpolarization-activated K+ channel KAT1 and a Xenopus depolarization-activated K+ channel. We report here that two of the chimeric constructs, which contain the first third of the KAT1 sequence, including the first four membrane-spanning segments (S1-S4) and the linker sequence between the fourth and fifth membrane-spanning segments, express functional channels that retain activation by hyperpolarization, but not depolarization. These two chimeric channels are no longer selective for K+. The chimeras are selective for cations over anions and are permeable to Ca2+. Therefore, unlike animal hyperpolarization-activated K+ channels, in which the carboxyl terminus is important for inward rectification induced by Mg2+ and polyamine block, the plant KAT1 channel has its major determinants for inward rectification in the amino-terminal region, which ends at the end of the S4-S5 linker.