Naamidine A intensifies the phosphotransferase activity of extracellular signal-regulated kinases causing A-431 cells to arrest in G1.

The binding of epidermal growth factor to its receptor activates the mitogen-activated protein kinase pathway. This pathway has been identified as a vital link between membrane-bound Ras and nuclear events and, therefore, is a potential target for chemotherapeutic drugs. We reported previously that naamidine A (NA), an alkaloid from the calcareous sponge Leucetta chagosensis, potently inhibited epidermal growth factor-stimulated DNA synthesis. In this current study, we demonstrate that in addition to its antimitogenic effects (complete inhibition of DNA synthesis at 0.78 micro M in A-431 cells after 30 h), NA at 1.56 micro M caused cells to arrest in the G(1) phase of the cell cycle. In vitro kinase, in-gel kinase, and Western blotting experiments demonstrate that extracellular signal-regulated kinase (ERK) 1 and ERK2 are primary molecular targets for NA in A-431 cells. Treatment with NA at concentrations between 0.78 and 3.13 micro M produces changes in the phosphorylation states of the ERKs, and strongly induces the phosphotransferase activity of the ERK enzymes. Our data indicate that treatment with NA generates a robust ERK signal. NA is the first small molecule reported to cause this effect on the ERK kinases and consequent G(1) cell cycle arrest.