The long myosin light chain kinase is differentially phosphorylated during interphase and mitosis.
We have shown previously that the activity of the long myosin light chain kinase (MLCK) is cell cycle regulated with a decrease in specific activity during mitosis that can be restored following treatment with alkaline phosphatase. To better understand the role and significance of phosphorylation in regulating MLCK function during mitosis, we examined the phosphorylation state of in vivo derived MLCK. Phosphoamino acid analysis and phosphopeptide mapping demonstrate that the long MLCK is differentially phosphorylated on serine residues during interphase and mitosis with the majority of the phosphorylation sites located within the N-terminal IgG domain. Biochemical assays show that Aurora B binds and phosphorylates the IgG domain of the long MLCK. In addition, phosphopeptide maps of the endogenous full-length MLCK from mitotic cells and in vitro phosphorylated IgG domain demonstrate that Aurora B phosphorylates the same sites as those observed in vivo. Altogether, these studies suggest that the long MLCK may be a cellular target for Aurora B during mitosis.
Dulyaninova
Natalya G
NG
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Bresnick
Anne R
AR
D000818:Animals; D064107:Aurora Kinase B; D064127:Aurora Kinases; D005982:Glutathione Transferase / Q000378:metabolism; D006367:HeLa Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007399:Interphase / Q000502:physiology; D008938:Mitosis / Q000502:physiology; D015879:Myosin Subfragments / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D010748:Phosphopeptides / Q000378:metabolism; D010766:Phosphorylation; D011233:Precipitin Tests; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011817:Rabbits; D012694:Serine / Q000737:chemistry / Q000235:genetics
D007074:Immunoglobulin G; D015879:Myosin Subfragments; D010446:Peptide Fragments; D010748:Phosphopeptides; D012694:Serine; D005982:Glutathione Transferase; C576921:AURKB protein, human; D064107:Aurora Kinase B; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases; D009219:Myosin-Light-Chain Kinase