The relationship between calcium, MAP kinase, and DNA synthesis in the sea urchin egg at fertilization.
Fertilization releases the brake on the cell cycle and the egg completes meiosis and enters into S phase of the mitotic cell cycle. The MAP kinase pathway has been implicated in this process, but the precise role of MAP kinase in meiosis and the first mitotic cell cycle remains unknown and may differ according to species. Unlike the eggs of most animals, sea urchin eggs have completed meiosis prior to fertilization and are arrested at the pronuclear stage. Using both phosphorylation-state-specific antibodies and a MAP kinase activity assay, we observe that MAP kinase is phosphorylated and active in unfertilized sea urchin eggs and then dephosphorylated and inactivated by 15 min postinsemination. Further, Ca(2+) was both sufficient and necessary for this MAP kinase inactivation. Treatment of eggs with the Ca(2+) ionophore A23187 caused MAP kinase inactivation and triggered DNA synthesis. When the rise in intracellular Ca(2+) was inhibited by injection of a chelator, BAPTA or EGTA, the activity of MAP kinase remained high. Finally, inhibition of the MAP kinase signaling pathway by the specific MEK inhibitor PD98059 triggered DNA synthesis in unfertilized eggs. Thus, whenever MAP kinase activity is retained, DNA synthesis is inhibited while inactivation of MAP kinase correlates with initiation of DNA synthesis.
Carroll
D J
DJ
Department of Molecular, Cellular and Developmental Biology and The Marine Science Institute, University of California at Santa Barbara, Santa Barbara, California, 93106-9610, USA.
Albay
D T
DT
Hoang
K M
KM
O'Neill
F J
FJ
Kumano
M
M
Foltz
K R
KR
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D004261:DNA Replication; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005306:Fertilization; D005419:Flavonoids / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010063:Ovum / Q000201:enzymology / Q000378:metabolism*; D010766:Phosphorylation; D012617:Sea Urchins; D015398:Signal Transduction
D004791:Enzyme Inhibitors; D005419:Flavonoids; D020928:Mitogen-Activated Protein Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D002118:Calcium