Mitosis-specific phosphorylation of smooth muscle regulatory light chain of myosin II at Ser-1 and/or -2 and Thr-9 in sea urchin egg extract.
We analyzed the kinase activities capable of phosphorylating the regulatory light chain of myosin-II (MRLC) from chicken gizzard in unfertilized and fertilized sea urchin egg extracts. Total kinase activity phosphorylating MRLC in vitro did not fluctuate throughout the first cell cycle. Phosphopeptide mapping analysis showed that MRLC was phosphorylated at two different sites corresponding to myosin light chain purified from chicken gizzard (MLCK) and protein kinase C (PKC) phosphorylation sites, namely MLCK and PKC sites, respectively. The activity of the kinase(s) responsible for phosphorylation of MRLC at PKC sites showed a significant increase at metaphase. Phosphoamino acid analysis revealed that this increase in MRLC phosphorylation was due to phosphorylation at serine residue (Ser-1 and/or Ser-2) and a threonine residue (Thr-9). This increase in phosphorylation at PKC sites is occurred concomitantly with an increase in histone H1 kinase activity. In contrast, MRLC phosphorylation at MLCK sites showed no significant changes during the first cell cycle. Butyrolactone I, a selective inhibitor of p34cdc2 kinase, inhibited the activity of the kinase(s) responsible for phosphorylation of MRLC at PKC sites at metaphase. These results suggest that the increase in MRLC phosphorylation at PKC sites (Ser-1 and/or -2, and Thr-9) at metaphase may be induced by p34cdc2 kinase. Thus, p34cdc2 kinase may be involved in the regulation of MRLC phosphorylation during cell division.
Totsukawa
G
G
Department of Biological Science, Faculty of Science, Hiroshima University, Japan.
Himi-Nakamura
E
E
Komatsu
S
S
Iwata
K
K
Tezuka
A
A
Sakai
H
H
Yazaki
K
K
Hosoya
H
H
D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D002453:Cell Cycle; D002645:Chickens; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005895:Gizzard, Avian; D008677:Metaphase; D008938:Mitosis; D009130:Muscle, Smooth / Q000378:metabolism*; D018994:Myosin Light Chains / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism*; D010063:Ovum; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D012617:Sea Urchins; D012694:Serine; D013912:Threonine
D004791:Enzyme Inhibitors; D018994:Myosin Light Chains; D013912:Threonine; D012694:Serine; C082868:butyrolactone I; D011493:Protein Kinase C; D009219:Myosin-Light-Chain Kinase; D015107:4-Butyrolactone