Chemical genetic analysis of the regulatory role of Cdc2p in the S. pombe septation initiation network.
The protein kinase Cdc2p is the master regulator of cell cycle progression in the fission yeast Schizosaccharomyces pombe. It is required both for entry into mitosis and for onset of DNA replication. Cdc2p must be inactivated to permit exit from mitosis, licensing of replication origins and cytokinesis. To study the role of Cdc2p in greater detail, we generated a cdc2 allele that is sensitive to an inhibitory ATP analogue. We show that the inhibitor-induced cell cycle arrest is reversible and examine the effect of inhibiting Cdc2p on the regulation of the septation initiation network (SIN), which controls the initiation of cytokinesis in S. pombe. We found that specific inactivation of Cdc2p in a mitotically arrested cell promotes the asymmetrical recruitment of SIN proteins to the spindle poles and the recruitment of the most downstream SIN components and beta-(1,3) glucan synthase to the contractile ring. Thus, we conclude that inactivation of Cdc2p is sufficient to activate the SIN and promote cytokinesis.
Dischinger
Sandra
S
Cell cycle control laboratory, ISREC, 1066 Epalinges, Switzerland.
Krapp
Andrea
A
Xie
Linfeng
L
Paulson
James R
JR
Simanis
Viesturs
V
D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D002453:Cell Cycle / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000502:physiology; D048749:Cytokinesis; D008540:Meiosis; D008938:Mitosis; D009154:Mutation; D011494:Protein Kinases / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000502:physiology; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D012568:Schizosaccharomyces / Q000166:cytology* / Q000201:enzymology / Q000235:genetics; D029702:Schizosaccharomyces pombe Proteins / Q000032:analysis / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*
C530021:4-amino-1-tert-butyl-3-(1'-naphthylmethyl)pyrazolo(3,4-d)pyrimidine; D018797:Cell Cycle Proteins; D011720:Pyrazoles; D011743:Pyrimidines; D029702:Schizosaccharomyces pombe Proteins; D011494:Protein Kinases; C409036:Sid1 kinase; D016203:CDC2 Protein Kinase; C469447:cdc2 protein, S pombe; C439269:Clp1 protein, S. pombe; D017027:Protein Tyrosine Phosphatases