Both Binding and Activation of p38 Mitogen-Activated Protein Kinase (MAPK) Play Essential Roles in Regulation of the Nucleocytoplasmic Distribution of MAPK-Activated Protein Kinase 5 by Cellular Stress

Abstract

The p38 mitogen-activated protein kinase (MAPK) pathway is an important mediator of cellular responses to environmental stress. Targets of p38 include transcription factors, components of the translational machinery, and downstream serine/threonine kinases, including MAPK-activated protein kinase 5 (MK5). Here we have used enhanced green fluorescent protein fusion proteins to analyze the subcellular localization of MK5. Although this protein is predominantly nuclear in unstimulated cells, MK5 shuttles between the nucleus and the cytoplasm. Furthermore, we have shown that the C-terminal domain of MK5 contains both a functional nuclear localization signal (NLS) and a leucine-rich nuclear export signal (NES), indicating that the subcellular distribution of this kinase reflects the relative activities of these two signals. In support of this, we have shown that stress-induced activation of the p38 MAPK stimulates the chromosomal region maintenance 1 protein-dependent nuclear export of MK5. This is regulated by both binding of p38 MAPK to MK5, which masks the functional NLS, and stress-induced phosphorylation of MK5 by p38 MAPK, which either activates or unmasks the NES. These properties may define the ability of MK5 to differentially phosphorylate both nuclear and cytoplasmic targets or alternatively reflect a mechanism whereby signals initiated by activation of MK5 in the nucleus may be transmitted to the cytoplasm.

We thank R. Davis, J. Han, and M. Gaestel for providing plasmids; Chris Armstrong (MRC Phosphorylation Unit, Dundee, Scotland) for PRAKtide, recombinant MK5(PRAK), anti-MK5(PRAK) antibodies, and purified GST-p38α; and Espen Michaelsen (University of Tromsø) for technical assistance.

This work was supported by grants from the Norwegian Cancer Society (DNK, project number A01037) and the Norwegian Research Council (project number 123686/310 and 135823/310), by the Erna and Olav Aakre Foundation for Fighting Cancer, and by Cancer Research UK. B.J. is supported by the Norwegian Cancer Society (DNK).