Dex-ras1 and serum- and glucocorticoid-inducible protein kinase 1: regulation of expression by dexamethasone in HEK293 cells.
The molecular and cellular basis of the psychotropic actions of adrenal corticosteroids is poorly understood. Previously, we reported that modulation of large conductance Ca2+-activated potassium channel (BK-channel) function by glucocorticoids can be recapitulated in human embryonic kidney293 (HEK293) cells (J Physiol 537:57, 2001). In the present paper, we examined the effect of dexamethasone on the expression of candidate mediator proteins of glucocorticoid action, dex-ras1 and serum and glucocorticoid inducible protein kinase 1 (SGK), in HEK293 cells. Dex-ras1 mRNA was readily detectable under basal conditions however, no changes of dex-ras1 mRNA expression occurred upon exposure to 100 nM of dexamethasone for 2 h. In contrast, a 2.5-fold increase of SGK mRNA was found under similar conditions. Total levels of cellular SGK protein were unaltered upon exposure to dexamethasone, but a marked increase of SGK in a Triton-X100 insoluble fraction was observed. BK-channel alpha-subunits could not be co-immunoprecipitated with SGK. In summary, SGK, but not dex-ras1, mRNA is rapidly induced by glucocorticoid stimulation in HEK293 cells. However, there appears to be no direct protein-protein interaction between SGK and BK-channel alpha-subunits.
Attarzadeh-Yazdi
Ghassem
G
Membrane Biology Group, Centre for Integrative Physiology, University of Edinburgh, Edinburgh, EH8 9XD, UK. gattarzadeh@yahoo.com
Shipston
Michael J
MJ
Antoni
Ferenc A
FA
D015152:Blotting, Northern; D045744:Cell Line, Tumor; D003907:Dexamethasone / Q000494:pharmacology*; D017930:Genes, Reporter / Q000187:drug effects / Q000235:genetics; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics*; D047468:Immunoprecipitation; D008156:Luciferases / Q000235:genetics; D024681:Potassium Channels, Calcium-Activated / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018631:ras Proteins / Q000235:genetics*; D000099184:Serum-Glucocorticoid Regulated Kinases
D003907:Dexamethasone; D017874:Immediate-Early Proteins; D008156:Luciferases; D024681:Potassium Channels, Calcium-Activated; D017346:Protein Serine-Threonine Kinases; D012333:RNA, Messenger; D018631:ras Proteins; D000099184:Serum-Glucocorticoid Regulated Kinases; C400132:RASD1 protein, human