Requirement for p34cdc2 kinase is restricted to mitosis in the mammalian cdc2 mutant FT210.
The mouse FT210 cell line is a temperature-sensitive cdc2 mutant. FT210 cells are found to arrest specifically in G2 phase and unlike many alleles of cdc2 and cdc28 mutants of yeasts, loss of p34cdc2 at the nonpermissive temperature has no apparent effect on cell cycle progression through the G1 and S phases of the division cycle. FT210 cells and the parent wild-type FM3A cell line each possess at least three distinct histone H1 kinases. H1 kinase activities in chromatography fractions were identified using a synthetic peptide substrate containing the consensus phosphorylation site of histone H1 and the kinase subunit compositions were determined immunochemically with antisera prepared against the "PSTAIR" peptide, the COOH-terminus of mammalian p34cdc2 and the human cyclins A and B1. The results show that p34cdc2 forms two separate complexes with cyclin A and with cyclin B1, both of which exhibit thermal lability at the non-permissive temperature in vitro and in vivo. A third H1 kinase with stable activity at the nonpermissive temperature is comprised of cyclin A and a cdc2-like 34-kD subunit, which is immunoreactive with anti-"PSTAIR" antiserum but is not recognized with antiserum specific for the COOH-terminus of p34cdc2. The cyclin A-associated kinases are active during S and G2 phases and earlier in the division cycle than the p34cdc2-cyclin B1 kinase. We show that mouse cells possess at least two cdc2-related gene products which form cell cycle regulated histone H1 kinases and we propose that the murine homolog of yeast p34cdc/CDC28 is essential only during the G2-to-M transition in FT210 cells.
Hamaguchi
J R
JR
Department of Biological Chemistry, School of Medicine, University of California, Davis 95616.
Tobey
R A
RA
Pines
J
J
Crissman
H A
HA
Hunter
T
T
Bradbury
E M
EM
D000595:Amino Acid Sequence; D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D002460:Cell Line; D002845:Chromatography; D016213:Cyclins / Q000502:physiology; D015151:Immunoblotting; D007399:Interphase; D016200:Maturation-Promoting Factor / Q000502:physiology; D051379:Mice; D008938:Mitosis / Q000502:physiology*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010446:Peptide Fragments; D010455:Peptides / Q000502:physiology; D016196:S Phase; D013696:Temperature
D016213:Cyclins; D010446:Peptide Fragments; D010455:Peptides; C062418:PSTAIR peptide; D016203:CDC2 Protein Kinase; D016200:Maturation-Promoting Factor