Cell cycle-dependent phosphorylation of smooth muscle myosin light chain in sea urchin egg extracts.
We studied enzymatic activities in sea urchin egg extracts that phosphorylate myosin regulatory light chain (MRLC) from chicken gizzard smooth muscle. The activity in the presence of EGTA showed cell cycle-dependent changes similar to that of histone H1 kinase, namely, it peaked shortly before cleavage, while that in the presence of Ca2+ ions did not show significant change during division cycle. Phosphopeptide mapping revealed that both the sites phosphorylatable by smooth muscle myosin light chain kinase (MLCK sites) and the sites phosphorylatable by protein kinase C (PKC sites) were phosphorylated in the presence or absence of Ca2+ ions. By analyses using an inhibitor of cdc2 kinase, butyrolactone-I, and ion exchange column chromatography, at least three kinases were detected as kinases that phosphorylate MRLC in vitro. These kinases phosphorylated distinct sites on MRLC. The first one, which phosphorylated the PKC sites, was identified as cdc2 kinase. The second one phosphorylated the MLCK sites in the absence of Ca2+ ions. The third one phosphorylated unknown sites. Possible implication of these activities in regulation of cytokinesis is discussed.
Mishima
M
M
Department of Biology, School of Arts and Sciences, University of Tokyo.
Mabuchi
I
I
D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002453:Cell Cycle; D002457:Cell Extracts; D002645:Chickens; D002852:Chromatography, Ion Exchange; D004533:Egtazic Acid / Q000494:pharmacology; D016200:Maturation-Promoting Factor / Q000378:metabolism; D009130:Muscle, Smooth / Q000201:enzymology; D018994:Myosin Light Chains / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000737:chemistry / Q000378:metabolism*; D010063:Ovum / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D012617:Sea Urchins
D002457:Cell Extracts; D018994:Myosin Light Chains; D004533:Egtazic Acid; C082868:butyrolactone I; D011493:Protein Kinase C; D009219:Myosin-Light-Chain Kinase; D016203:CDC2 Protein Kinase; D016200:Maturation-Promoting Factor; D015107:4-Butyrolactone; D002118:Calcium