Tetramerization of the AKT1 plant potassium channel involves its C-terminal cytoplasmic domain.
All plant channels identified so far show high conservation throughout the polypeptide sequence except in the ankyrin domain which is present only in those closely related to AKT1. In this study, the architecture of the AKT1 protein has been investigated. AKT1 polypeptides expressed in the baculovirus/Sf9 cells system were found to assemble into tetramers as observed with animal Shaker-like potassium channel subunits. The AKT1 C-terminal intracytoplasmic region (downstream from the transmembrane domain) alone formed tetrameric structures when expressed in Sf9 cells, revealing a tetramerization process different from that of Shaker channels. Tests of subfragments from this sequence in the two-hybrid system detected two kinds of interaction. The first, involving two identical segments (amino acids 371-516), would form a contact between subunits, probably via their putative cyclic nucleotide-binding domains. The second interaction was found between the last 81 amino acids of the protein and a region lying between the channel hydrophobic core and the putative cyclic nucleotide-binding domain. As the interacting regions are highly conserved in all known plant potassium channels, the structural organization of AKT1 is likely to extend to these channels. The significance of this model with respect to animal cyclic nucleotide-gated channels is also discussed.
Daram
P
P
Laboratoire de Biochimie et Physiologie Moléculaire des Plantes, ENSA-M/INRA/CNRS URA 2133/UM2, Montpellier, France.
Urbach
S
S
Gaymard
F
F
Sentenac
H
H
Chérel
I
I
D000595:Amino Acid Sequence; D000818:Animals; D029681:Arabidopsis Proteins; D001665:Binding Sites; D002460:Cell Line; D003593:Cytoplasm; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015221:Potassium Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera / Q000166:cytology
D029681:Arabidopsis Proteins; D010940:Plant Proteins; D015221:Potassium Channels; D011993:Recombinant Fusion Proteins; C074500:AKT1 protein, Arabidopsis