DNA replication is completed in Saccharomyces cerevisiae cells that lack functional Cdc14, a dual-specificity protein phosphatase

Abstract The Cdc14 protein encodes a dual-specificity protein phosphatase which functions in late mitosis, and considerable genetic evidence suggests a role in DNA replication. We find that cdc14 mutants arrested in late mitosis maintain persistent levels of mitotic kinase activity, suggesting that Cdc14 controls inactivation of this kinase. Overexpression of Sic1, a cyclin-dependent protein kinase inhibitor, is able to suppress telophase mutants such as dbf2, cdc5 and cdc15, but not cdc14. It does, however, force cdc14-arrested cells into the next cell cycle, in which an apparently normal S phase occurs as judged by FACS and pulsed-field gel electrophoretic analysis. Furthermore, in a promoter shut-off experiment, cells lacking Cdc14 appear to carry out a normal S phase. Thus Cdc14 functions mainly in late mitosis and it has no essential role in S phase.

[1]  L. Johnston,et al.  Spo12 is a limiting factor that interacts with the cell cycle protein kinases Dbf2 and Dbf20, which are involved in mitotic chromatid disjunction. , 1993, Genetics.

[2]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[3]  D. Koshland,et al.  Mitotic transmission of artificial chromosomes in cdc mutants of the yeast, Saccharomyces cerevisiae. , 1990, Genetics.

[4]  L. Johnston,et al.  The Swi5 transcription factor of Saccharomyces cerevisiae has a role in exit from mitosis through induction of the cdk-inhibitor Sic1 in telophase. , 1997, Genetics.

[5]  L. Hartwell,et al.  Genetic control of the cell division cycle in yeast. 3. Seven genes controlling nuclear division. , 1971, Experimental cell research.

[6]  G. Taylor,et al.  The Activity of Cdc 14 p , an Oligomeric Dual Specificity Protein Phosphatase from Saccharomyces cerevisiae , Is Required for Cell Cycle Progression , 1997 .

[7]  L. Hartwell,et al.  Genetic Control of the Cell Division Cycle in Yeast: V. Genetic Analysis of cdc Mutants. , 1973, Genetics.

[8]  P. Hieter,et al.  Establishing genetic interactions by a synthetic dosage lethality phenotype. , 1996, Genetics.

[9]  M. Mendenhall,et al.  An inhibitor of p34CDC28 protein kinase activity from Saccharomyces cerevisiae. , 1993, Science.

[10]  D. Botstein,et al.  A group of interacting yeast DNA replication genes. , 1991, Genes & Development.

[11]  M. Shirayama,et al.  Dominant mutant alleles of yeast protein kinase gene , 1996 .

[12]  C. F. Hardy.  Characterization of an essential Orc2p-associated factor that plays a role in DNA replication , 1996, Molecular and cellular biology.

[13]  Bruce Stillman,et al.  ORC and Cdc6p interact and determine the frequency of initiation of DNA replication in the genome , 1995, Cell.

[14]  L. Johnston,et al.  P40SDB25, a putative CDK inhibitor, has a role in the M/G1 transition in Saccharomyces cerevisiae. , 1994, Genes & development.

[15]  L. Hartwell,et al.  Altered fidelity of mitotic chromosome transmission in cell cycle mutants of S. cerevisiae. , 1985, Genetics.

[16]  A. Toh-E,et al.  The yeast TEM1 gene, which encodes a GTP-binding protein, is involved in termination of M phase , 1994, Molecular and cellular biology.

[17]  R. Deshaies.  Phosphorylation and proteolysis: partners in the regulation of cell division in budding yeast. , 1997, Current opinion in genetics & development.

[18]  R. D. Gietz,et al.  New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. , 1988, Gene.