Regulatory differences between atypical and typical MAP kinases in Dictyostelium discoideum.

Within the large family of mitogen activated protein kinases (MAPKs), one outlier group referred to as atypical MAPKs is not regulated by conventional upstream MAPK kinases (MAP2Ks). This includes the Dictyostelium discoideum atypical MAPK Erk2, a protein kinase essential for chemotactic movement and development. The regulation and functional specificity of Erk2 was investigated through phenotypic analysis of chimeric and mutant MAPKs. Chimeric MAPKs containing regions of Erk2 were created using complementary regions of the more typical MAPK Erk1, that provides very different functions in this amoeba. The chimeric MAPKs were not phosphorylated at levels observed for wild-type MAPKs and none rescued wild-type MAPK function to erk1 or erk2 cells. Endogenous Erk1 and Erk2 MAPKs were destabilized in cells expressing chimeric MAPKs containing the same carboxyl terminus. A carboxyl terminal motif conserved among atypical MAPKs was important but not essential for Erk2 regulation and function and the motif did not confer atypical MAPK regulation when present in Erk1. A kinase-dead version of Erk2 was phosphorylated in response to folate or cAMP chemotactic stimulation, suggesting Erk2 is activated in vivo by an upstream protein kinase, contrary to previous predictions of autophosphorylation. This regulation implies a protein kinase distinct from the single conventional MAP2K in Dictyostelium regulates atypical MAPK signaling. A non-activatable form of Erk2 was not capable of rescuing Erk2 function in erk2 cells. These findings suggest that the regulation of atypical and typical MAPKs is substantially different and carried out by distinct upstream protein kinases.