The stimulus-dependent co-localization of serum- and glucocorticoid-regulated protein kinase (Sgk) and Erk/MAPK in mammary tumor cells involves the mutual interaction with the importin-alpha nuclear import protein.

Abstract

In Con8 rat mammary epithelial tumor cells, indirect immunofluorescence revealed that Sgk (serum- and glucocorticoid-regulated kinase) and Erk/MAPK (extracellular signal-regulated protein kinase/mitogen activated protein kinase) co-localized to the nucleus in serum-treated cells and to the cytoplasmic compartment in cells treated with the synthetic glucocorticoid dexamethasone. Moreover, the subcellular distribution of the importin-alpha nuclear transport protein was similarly regulated in a signal-dependent manner. In vitro GST-pull down assays revealed the direct interaction of importin-alpha with either Sgk or Erk/MAPK, while RNA interference knockdown of importin-alpha expression disrupted the localization of both Sgk and Erk into the nucleus of serum-treated cells. Wild type or kinase dead forms of Sgk co-immunoprecipitated with Erk/MAPK from either serum- or dexamethasone-treated mammary tumor cells, suggesting the existence of a protein complex containing both kinases. In serum-treated cells, nucleus residing Sgk and Erk/MAPK were both hyperphosphorylated, indicative of their active states, whereas, in dexamethasone-treated cells Erk/MAPK, but not Sgk, was in its inactive hypophosphorylated state. Treatment with a MEK inhibitor, which inactivates Erk/MAPK, caused the relocalization of both Sgk and ERK to the cytoplasm. We therefore propose that the signal-dependent co-localization of Sgk and Erk/MAPK mediated by importin-alpha represents a new pathway of signal integration between steroid and serum/growth factor-regulated pathways.

• Authors: Patricia Buse, Anita C Maiyar, Kim L Failor, Susan Tran, Meredith L L Leong, Gary L Firestone
• Journal: Experimental cell research (Exp Cell Res) Vol. 313 Issue 15 Pg. 3261-75 (Sep 10 2007) ISSN: 0014-4827 [Print] United States
• PMID: 17692313 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Glucocorticoids
• Immediate-Early Proteins
• Nuclear Localization Signals
• alpha Karyopherins
• Dexamethasone
• Protein Serine-Threonine Kinases
• serum-glucocorticoid regulated kinase
• Extracellular Signal-Regulated MAP Kinases

Topics

• Active Transport, Cell Nucleus
• Animals
• Cell Line, Tumor
• Cell Nucleus (metabolism)
• Cytoplasm (metabolism)
• Dexamethasone (pharmacology)
• Extracellular Signal-Regulated MAP Kinases (metabolism)
• Glucocorticoids (pharmacology)
• Immediate-Early Proteins (metabolism)
• Mammary Neoplasms, Animal
• Nuclear Localization Signals (metabolism)
• Phosphorylation
• Protein Binding
• Protein Serine-Threonine Kinases (metabolism)
• Rats
• alpha Karyopherins (metabolism)