A more complex, specific example of crosstalk between two major signaling pathways can be observed with the interaction of the cAMP and MAPK signaling pathways in the activation of lymphocytes. In this case, components of the cAMP pathway directly and indirectly affect MAPK signaling pathway meant to activate genes involving immunity and lymphocytes.
Newly formed cAMP is released from the membrane and diffuses across the intracellular space where it serves to activate PKA. The catalytic subunit of PKA must bind four molecules of cAMP to be activated, whereupon activation consists of cleavage between the regulatory and catalytic subunits. This cleavage in turn activates PKA by exposing the catalytic sites of the C subunits, which can then phosphorylate an array of proteins in the cell.
In lymphocytes, the intracellular levels of cAMP increase upon antigen-receptor stimulation and even more so in response to prostaglandin E and other immunosuppression agents. In this case, cAMP serves to inhibit immunity players. PKA type I colocalizes with the T-cell and B-cell antigen receptors and causes inhibition of T- and B-cell activation. PKA has even been highlighted as a direct inducer of genes contributing to immunosuppression.
Additionally, the cAMP pathway also interacts with the MAPK pathway in a more indirect manner through its interaction with hematopoietic PTPase (HePTP). HePTP is expressed in all leukocytes. When overexpressed in T-cells, HePTP reduces the transcriptional activation of the interleukin-2 promoter typically induced by the activated T-cell receptor through a MAPK signaling cascade. The way that HePTP effectively inhibits the MAPK signaling is by interacting with the MAP kinases Erk1, Erk2, and p38 through a short sequence in HePTP's non-catalytic N terminus termed the kinase interaction motif (KIM)., The highly-specific binding of Erk and p38 to this subunit of HePTP results in rapid inactivation of the signaling cascade (see figure 3).
Yet, since both HePTP and Erk are cytosolic enzymes, it is reasonable to conclude that there exists a mechanism for the inhibition of Erk by HePTP to cease in order to allow for the translocation of activated Erk to the nucleus. Indeed, like in many other cases of protein-protein interaction, HePTP appears to be phosphorylated by Erk and p38 at the sites Thr45 and Ser72. Importantly though, a third phosphorylation site in the non-catalytic N terminus (the KIM region) of HePTP has been found—one that is phosphorylated to a much higher stoichiometry by the cAMP pathway, in yet another instance of crosstalk between the cAMP and MAPK pathways.
Phosphorylation of this third site by PKAs from the cAMP pathway inhibits binding of MAP kinases to HePTP and thereby upregulates the MAPK/ERK signaling cascade. The MAPK pathway, through Ras, Raf, Mek, and Erk, shows low activity in the presence of unphosphorylated (active) HePTP. However, activation the cAMP pathway stimulates the activation of PKA, which in turn phosphorylates HePTP at Ser23. This prevents HePTP from binding to Erk and frees the MAPK pathway from inhibition, allowing downstream signaling to continue (see figure 4).
Moreover, studies involving smooth muscle cells from the atrium of the heart have shown that PKA can reduce the activation of MAP kinases in response to platelet-derived growth factor (PDGF) by phosphorylating the kinase c-Raf. Thus, it seems plausible that PKA in the cAMP pathway could even be further involved in the regulation of lymphocyte activation not only by inhibiting the antigen-receptor MAPK signal pathway at its final stage, but even further upstream.