Functions of AKT1 and AKT2 potassium channels determined by studies of single and double mutants of Arabidopsis.
A reverse genetic strategy was used to isolate Arabidopsis plants containing "knockout" mutations in AKT1 and AKT2, two members of a K+ channel gene family. Comparative studies of growth and membrane properties in wild-type and mutant seedlings were performed to investigate the physiological functions of these two related channels. The growth rates of plants supplied with rate-limiting concentrations of K+ depended on the presence of AKT1 but not AKT2 channels. This result indicates that AKT1 but not AKT2 mediates growth-sustaining uptake of K+ into roots, consistent with the expression patterns of these two genes. K+ -induced membrane depolarizations were measured with microelectrodes to assess the contribution each channel makes to the K+ permeability of the plasma membrane in three different organs. In apical root cells, AKT1 but not AKT2 contributed to the K+ permeability of the plasma membrane. In cotyledons, AKT1 was also the principal contributor to the K+ permeability. However, in the mesophyll cells of leaves, AKT2 accounted for approximately 50% of the K+ permeability, whereas AKT1 unexpectedly accounted for the remainder. The approximately equal contributions of AKT1 and AKT2 in leaves detected by the in vivo functional assay employed here are not in agreement with previous RNA blots and promoter activity studies, which showed AKT2 expression to be much higher than AKT1 expression in leaves. This work demonstrates that comparative functional studies of specific mutants can quantify the relative contributions of particular members of a gene family, and that expression studies alone may not reliably map out distribution of gene functions.
Dennison
K L
KL
Biotechnology Center, University of Wisconsin, 425 Henry Mall, Madison, WI 53706, USA.
Robertson
W R
WR
Lewis
B D
BD
Hirsch
R E
RE
Sussman
M R
MR
Spalding
E P
EP
D017360:Arabidopsis / Q000235:genetics / Q000502:physiology*; D029681:Arabidopsis Proteins; D002463:Cell Membrane Permeability / Q000235:genetics / Q000502:physiology; D018548:Cotyledon / Q000235:genetics / Q000254:growth & development; D004251:DNA Transposable Elements; D018506:Gene Expression Regulation, Plant; D009154:Mutation; D010641:Phenotype; D010940:Plant Proteins / Q000235:genetics / Q000502:physiology*; D018517:Plant Roots / Q000235:genetics / Q000502:physiology; D011188:Potassium / Q000378:metabolism*; D015221:Potassium Channels / Q000235:genetics / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt
D029681:Arabidopsis Proteins; D004251:DNA Transposable Elements; D010940:Plant Proteins; D015221:Potassium Channels; D011518:Proto-Oncogene Proteins; C074500:AKT1 protein, Arabidopsis; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D011188:Potassium