Co-expression of calcium-dependent protein kinase with the inward rectified guard cell K+ channel KAT1 alters current parameters in Xenopus laevis oocytes.
Increased guard cell cytosolic [Ca2+] is known to be involved in signal transduction pathways leading to stomatal closure, and inhibit the inward rectifying guard cell K+ channel KAT1. Guard cell calcium-dependent protein kinase (CDPK) has been shown to phosphorylate KAT1; such phosphorylation is known to modulate other K+ channels involved in signal transduction cascades. The work reported here focused on demonstrating CDPK-dependent inhibition of KAT1 currents. A cDNA encoding soybean CDPK was generated and it's translation product was shown to be functional; demonstrating Ca2+-dependent autophosphorylation and phosphorylation of a target protein. Ion currents were monitored using voltage clamp techniques upon expression of KAT1 in Xenopus laevis oocytes. Coexpression of recombinant CDPK with KAT1 in oocytes altered the kinetics and magnitude of induced K+ currents; at a given hyperpolarizing command voltage, the magnitude of KAT1 currents was reduced and the half-time for channel activation was increased. This finding supports a model of Ca2+-dependent ABA inhibition of inward K+ currents in guard cells as being mediated by CDPK phosphorylation of KAT1.
Berkowitz
G
G
Department of Plant Science U-67, University of Connecticut, Storrs 06269-4067, USA. gerald.berkowitz@uconn.edu
Zhang
X
X
Mercie
R
R
Leng
Q
Q
Lawton
M
M
D000818:Animals; D002118:Calcium / Q000378:metabolism; D003001:Cloning, Molecular; D003600:Cytosol / Q000378:metabolism; D004926:Escherichia coli; D005260:Female; D007700:Kinetics; D008564:Membrane Potentials / Q000502:physiology*; D009865:Oocytes / Q000502:physiology*; D010940:Plant Proteins / Q000235:genetics / Q000502:physiology; D015221:Potassium Channels / Q000235:genetics / Q000502:physiology*; D024661:Potassium Channels, Inwardly Rectifying; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D018075:RNA, Complementary; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction; D014982:Xenopus laevis; D017871:Calcium-Calmodulin-Dependent Protein Kinases
D002118:Calcium; D010940:Plant Proteins; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D011494:Protein Kinases; D018075:RNA, Complementary; D011994:Recombinant Proteins; D017871:Calcium-Calmodulin-Dependent Protein Kinases