MAP kinases are central signaling proteins that induce and regulate proliferation, differentiation, survival, and stress response. The predominant localization of the MAP kinases in quiescent cells is the cytoplasm, which is mostly due to their interaction with anchoring proteins. Stimulation releases the MAP kinase molecules from the cytoplasmic anchors, which allows their rapid nuclear translocation and regulation of gene transcription, which is important for the induction of the various processes. We found that the translocation of MAPKs is not mediated by the canonical NLS/importina/importinb mechanism, but rather involves b-like importins that act in at least two distinct manners to execute their function. ERK1/2 translocation is first mediated by the phosphorylation of their activatory residues, which allows their release from anchoring proteins. This is followed by CK2-mediated phosphorylation of two Ser residues in a nuclear translocation signal (NTS), that induces the binding of ERK1/2 to importin7 and penetration to the nucleus, via the nuclear pores. Despite the pronounced sequence and conformation similarity of JNK and p38 to the ERK1/2, we found that they use a different mechanism for their stimulated translocation. Thus, they can translocate to the nucleus in an activatory phosphorylation-independent manner, which most likely involves phosphorylation of their anchoring proteins. This is followed by the binding of JNK and p38 to either importin7 or importin9, each within dimers with Imp3, which translocate to the nuclear envelope and mediate the penetration of these MAPKs via the nuclear pores. This elucidation of the novel mechanism of MAP kinase translocation can be used to develop translocation inhibitors for combating MAP kinases-mediated pathologies.