Subcellular location of serum- and glucocorticoid-induced kinase-1 in renal and mammary epithelial cells.
Serum- and glucocorticoid-induced kinase-1 (SGK1) is involved in aldosterone-induced Na(+) reabsorption by increasing epithelial Na(+) channel (ENaC) activity in cortical collecting duct (CCD) cells, but its exact mechanisms of action are unknown. Although several potential targets such as Nedd4-2 have been described in expression systems, endogenous substrates mediating SGK1's physiological effects remain to be identified. In addition, subcellular localization studies of SGK1 have provided controversial results. We determined the subcellular location of SGK1 using SGK1-autofluorescent protein (AFP) fusion proteins. Rabbit CCD (RCCT-28A) cells were transiently transfected with a construct encoding for SGK1-AFP and were stained or cotransfected with markers for various subcellular compartments. In live cells, transiently expressed SGK1-AFP clearly colocalized with the mitochondrial marker rhodamine 123. Similarly, SGK1-AFP colocalized with the mitochondrial marker MitoTracker when stably expressed using a retroviral system in either RCCT-28A cells or the mammary epithelial cell line MCF10A. To determine which region of SGK1 is responsible for this subcellular localization, we generated RCCT-28A cell lines stably expressing SGK1 mutants. The results indicate that the NH(2)-terminal 60-amino acid region of SGK1 is necessary and sufficient for its subcellular localization. Localization of SGK1 to the mitochondria raises the possibility that SGK1 may play a role in regulating energy metabolism.
Cordas
Emily
E
Department of Physiology, Dartmouth Medical School, 1 Medical Center Dr., Lebanon, NH 03756, USA.
Náray-Fejes-Tóth
Anikó
A
Fejes-Tóth
Géza
G
D000818:Animals; D002460:Cell Line; D004847:Epithelial Cells / Q000201:enzymology*; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D017874:Immediate-Early Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000201:enzymology*; D042361:Mammary Glands, Human / Q000166:cytology / Q000201:enzymology*; D051379:Mice; D008928:Mitochondria / Q000201:enzymology*; D009154:Mutation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014162:Transfection; D000099184:Serum-Glucocorticoid Regulated Kinases
D049452:Green Fluorescent Proteins; D017874:Immediate-Early Proteins; D017346:Protein Serine-Threonine Kinases; D011993:Recombinant Fusion Proteins; D000099184:Serum-Glucocorticoid Regulated Kinases