p13suc1 suppresses the catalytic function of p34cdc2 kinase for intermediate filament proteins, in vitro.
The regulation of p34cdc2 kinase activity controls the entry into and exit from mitosis. Although genetic and biochemical evidence suggested close interactions between cyclins, p13suc1 and p34cdc2 kinase, the roles of p13suc1 on p34cdc2 kinase functions remain unclear. To examine the effects of p13suc1 on p34cdc2 kinase function we developed a simple purification procedure for p34cdc2 kinase, unassociated with p13suc1. The key to the purification procedures we used was buffer containing 0.5 M NaCl and 50% ethylene glycol, as a specific elutant of p34cdc2 kinase from p13suc1-Sepharose. This purified p34cdc2 kinase stoichiometrically phosphorylated vimentin and desmin. Exogenous p13suc1 suppressed the phosphorylation of these filament proteins by the kinase and prevented disassembly, although histone H1 phosphorylation was not affected. Peptide mapping analysis showed a similar extent of inhibition by p13suc1 for all five phosphorylation sites by p34cdc2 kinase of vimentin and desmin, hence these p13suc1-induced inhibitions are probably not site-specific. It thus appears that p13suc1 has a selective effect on the catalytic activity of p34cdc2 kinase for these filament proteins.
Kusubata
M
M
Department of Neurophysiology, Tokyo Metropolitan Institute of Gerontology, Japan.
Tokui
T
T
Matsuoka
Y
Y
Okumura
E
E
Tachibana
K
K
Hisanaga
S
S
Kishimoto
T
T
Yasuda
H
H
Kamijo
M
M
Ohba
Y
Y
D000595:Amino Acid Sequence; D000906:Antibodies; D015153:Blotting, Western; D016203:CDC2 Protein Kinase / Q000302:isolation & purification / Q000378:metabolism*; D018797:Cell Cycle Proteins; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002851:Chromatography, High Pressure Liquid; D003893:Desmin / Q000378:metabolism / Q000648:ultrastructure; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D007382:Intermediate Filaments / Q000378:metabolism / Q000648:ultrastructure; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D029702:Schizosaccharomyces pombe Proteins; D014746:Vimentin / Q000378:metabolism / Q000648:ultrastructure
D000906:Antibodies; D018797:Cell Cycle Proteins; D003893:Desmin; D005656:Fungal Proteins; D006657:Histones; D007381:Intermediate Filament Proteins; D010446:Peptide Fragments; D010455:Peptides; D011994:Recombinant Proteins; D029702:Schizosaccharomyces pombe Proteins; C051655:Suc1 protein, S pombe; D014746:Vimentin; D016203:CDC2 Protein Kinase