Early events in DNA replication require cyclin E and are blocked by p21CIP1

Using immunodepletion of cyclin E and the inhibitor protein p21WAF/CIP1, we demonstrate that the cyclin E protein, in association with Cdk2, is required for the elongation phase of replication on single-stranded substrates. Although cyclin E/Cdk2 is likely to be the major target by which p21 inhibits the initiation of sperm DNA replication, p21 can inhibit single-stranded replication through a mechanism dependent on PCNA. While the cyclin E/Cdk2 complex appears to have a role in the initiation of DNA replication, another Cdk kinase, possibly cyclin A/Cdk, may be involved in a later step controlling the switch from initiation to elongation. The provision of a large maternal pool of cyclin E protein shows that regulators of replication are constitutively present, which explains the lack of a protein synthesis requirement for replication in the early embryonic cell cycle.

[1]  T. Hunt,et al.  Identification of a developmental timer regulating the stability of embryonic cyclin A and a new somatic A-type cyclin at gastrulation. , 1995, Genes & development.

[2]  James M. Roberts,et al.  Human cyclin E, a nuclear protein essential for the G1-to-S phase transition , 1995, Molecular and cellular biology.

[3]  J. Newport,et al.  An analysis of the regulation of DNA synthesis by cdk2, Cip1, and licensing factor , 1995, The Journal of cell biology.

[4]  M. Kirschner,et al.  Separate domains of p21 involved in the inhibition of Cdk kinase and PCNA , 1995, Nature.

[5]  M. Kirschner,et al.  Mitosis in transition , 1994, Cell.

[6]  James M. Roberts,et al.  Rules to replicate by , 1994, Cell.

[7]  T. Hunter,et al.  Cyclins and cancer II: Cyclin D and CDK inhibitors come of age , 1994, Cell.

[8]  C. Sherr.  G1 phase progression: Cycling on cue , 1994, Cell.

[9]  U. Strausfeld,et al.  Cip1 blocks the initiation of DNA replication in Xenopus extracts by inhibition of cyclin-dependent kinases , 1994, Current Biology.

[10]  U. K. Laemmli,et al.  Study of the cell cycle‐dependent assembly of the DNA pre‐replication centres in Xenopus egg extracts. , 1994, The EMBO journal.

[11]  S. Elledge,et al.  Cdk-interacting protein 1 directly binds with proliferating cell nuclear antigen and inhibits DNA replication catalyzed by the DNA polymerase delta holoenzyme. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[12]  G. Hannon,et al.  p21-containing cyclin kinases exist in both active and inactive states. , 1994, Genes & development.

[13]  James M. Roberts,et al.  Cloning of p27 Kip1 , a cyclin-dependent kinase inhibitor and a potential mediator of extracellular antimitogenic signals , 1994, Cell.

[14]  Tony Hunter,et al.  p27, a novel inhibitor of G1 cyclin-Cdk protein kinase activity, is related to p21 , 1994, Cell.

[15]  Kim Nasmyth,et al.  Closing the cell cycle circle in yeast: G2 cyclin proteolysis initiated at mitosis persists until the activation of G1 cyclins in the next cycle , 1994, Cell.

[16]  G. Hannon,et al.  The p21 inhibitor of cyclin-dependent kinases controls DNA replication by interaction with PCNA , 1994, Nature.

[17]  C. Lehner,et al.  Cyclin E controls S phase progression and its down-regulation during Drosophila embryogenesis is required for the arrest of cell proliferation , 1994, Cell.

[18]  B. Stillman,et al.  Reconstitution of complete SV40 DNA replication with purified replication factors. , 1994, The Journal of biological chemistry.

[19]  M. Gossen,et al.  Acceleration of the G1/S phase transition by expression of cyclins D1 and E with an inducible system. , 1994, Molecular and cellular biology.

[20]  S. van den Heuvel,et al.  Distinct roles for cyclin-dependent kinases in cell cycle control. , 1993, Science.

[21]  David Beach,et al.  p21 is a universal inhibitor of cyclin kinases , 1993, Nature.

[22]  C. Turck,et al.  Inhibition of CDK2 activity in vivo by an associated 20K regulatory subunit , 1993, Nature.

[23]  S. Elledge,et al.  The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases , 1993, Cell.

[24]  J. Trent,et al.  WAF1, a potential mediator of p53 tumor suppression , 1993, Cell.

[25]  J. Newport,et al.  Distinct roles of cdk2 and cdc2 in RP-A phosphorylation during the cell cycle. , 1993, Journal of cell science.

[26]  H. Leonhardt,et al.  Reversal of terminal differentiation and control of DNA replication: Cyclin A and cdk2 specifically localize at subnuclear sites of DNA replication , 1993, Cell.

[27]  Charles J. Sherr,et al.  Mammalian G1 cyclins , 1993, Cell.

[28]  J. Pines,et al.  Cyclins and cyclin-dependent kinases: take your partners , 1993 .

[29]  F. Zindy,et al.  Localization of cyclin A at the sites of cellular DNA replication. , 1993, Experimental cell research.

[30]  James M. Roberts,et al.  Negative regulation of G1 in mammalian cells: inhibition of cyclin E-dependent kinase by TGF-beta. , 1993, Science.

[31]  R Pepperkok,et al.  Regulation of the cell cycle by the cdk2 protein kinase in cultured human fibroblasts , 1993, The Journal of cell biology.

[32]  James M. Roberts,et al.  Cyclin-dependent regulation of G1 in mammalian fibroblasts. , 1993, Science.

[33]  S. Elledge,et al.  Formation and activation of a cyclin E-cdk2 complex during the G1 phase of the human cell cycle. , 1992, Science.

[34]  M. Wigler,et al.  Oncogenic ras triggers the activation of 42-kDa mitogen-activated protein kinase in extracts of quiescent Xenopus oocytes. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[35]  S. Reed,et al.  Cyclin E/cdk2 and cyclin A/cdk2 kinases associate with p107 and E2F in a temporally distinct manner. , 1992, Genes & development.

[36]  U. K. Laemmli,et al.  Identification of nuclear pre-replication centers poised for DNA synthesis in Xenopus egg extracts: immunolocalization study of replication protein A , 1992, The Journal of cell biology.

[37]  S. Reed,et al.  Association of human cyclin E with a periodic G1-S phase protein kinase. , 1992, Science.

[38]  R. Weinberg,et al.  Regulation of retinoblastoma protein functions by ectopic expression of human cyclins , 1992, Cell.

[39]  R. Laskey,et al.  The nuclear membrane prevents replication of human G2 nuclei but not G1 nuclei in Xenopus egg extract , 1992, Cell.

[40]  R. Pepperkok,et al.  Cyclin A is required at two points in the human cell cycle. , 1992, The EMBO journal.

[41]  U. Strausfeld,et al.  Cyclin a is required for the onset of DNA replication in mammalian fibroblasts , 1991, Cell.

[42]  S. Reed,et al.  Isolation of three novel human cyclins by rescue of G1 cyclin (cln) function in yeast , 1991, Cell.

[43]  James M. Roberts,et al.  Human cyclin E, a new cyclin that interacts with two members of the CDC2 gene family , 1991, Cell.

[44]  J. Newport,et al.  Evidence that the G1-S and G2-M transitions are controlled by different cdc2 proteins in higher eukaryotes , 1991, Cell.

[45]  T. Hunt,et al.  On the synthesis and destruction of A- and B-type cyclins during oogenesis and meiotic maturation in Xenopus laevis , 1991, The Journal of cell biology.

[46]  T. Mitchison,et al.  Mitotic spindle assembly by two different pathways in vitro , 1991, The Journal of cell biology.

[47]  Marc W. Kirschner,et al.  Cyclin activation of p34 cdc2 , 1990, Cell.

[48]  P. Nurse,et al.  A cdc2-like protein is involved in the initiation of DNA replication in Xenopus egg extracts , 1990, Cell.

[49]  M. Méchali,et al.  Characterization and developmental expression of Xenopus proliferating cell nuclear antigen (PCNA). , 1990, Developmental biology.

[50]  T. Hunt,et al.  The A‐ and B‐type cyclin associated cdc2 kinases in Xenopus turn on and off at different times in the cell cycle. , 1990, The EMBO journal.

[51]  D. Newmeyer,et al.  An N-ethylmaleimide-sensitive cytosolic factor necessary for nuclear protein import: requirement in signal-mediated binding to the nuclear pore , 1990, The Journal of cell biology.

[52]  J. White,et al.  Replication occurs at discrete foci spaced throughout nuclei replicating in vitro. , 1989, Journal of cell science.

[53]  Tony Hunter,et al.  Isolation of a human cyclin cDNA: Evidence for cyclin mRNA and protein regulation in the cell cycle and for interaction with p34cdc2 , 1989, Cell.

[54]  I. Raška,et al.  Ultrastructural immunolocalization of cyclin/PCNA in synchronized 3T3 cells. , 1989, Experimental cell research.

[55]  D A Agard,et al.  Direct cell lineage analysis in Drosophila melanogaster by time-lapse, three-dimensional optical microscopy of living embryos , 1989, The Journal of cell biology.

[56]  E. Tan,et al.  Quantitation and subcellular localization of proliferating cell nuclear antigen (PCNA/cyclin) in oocytes and eggs of Xenopus laevis. , 1989, Experimental cell research.

[57]  Andrew W. Murray,et al.  Cyclin synthesis drives the early embryonic cell cycle , 1989, Nature.

[58]  C. Hutchison,et al.  DNA replication and cell cycle control in Xenopus egg extracts , 1989, Journal of Cell Science.

[59]  R Berezney,et al.  Mapping replicational sites in the eucaryotic cell nucleus , 1989, The Journal of cell biology.

[60]  J. Blow,et al.  A role for the nuclear envelope in controlling DNA replication within the cell cycle , 1988, Nature.

[61]  J. Blow,et al.  Steps in the assembly of replication-competent nuclei in a cell-free system from Xenopus eggs , 1988, The Journal of cell biology.

[62]  C. Hutchison,et al.  Periodic DNA synthesis in cell‐free extracts of Xenopus eggs. , 1987, The EMBO journal.

[63]  J. Blow,et al.  Nuclei act as independent and integrated units of replication in a Xenopus cell‐free DNA replication system. , 1987, The EMBO journal.

[64]  J. Newport,et al.  Disassembly of the nucleus in mitotic extracts: Membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes , 1987, Cell.

[65]  J. Blow,et al.  Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs , 1986, Cell.

[66]  J. Maller,et al.  Induction of nuclear envelope breakdown, chromosome condensation, and spindle formation in cell-free extracts , 1985, The Journal of cell biology.

[67]  D. Lane,et al.  An immunoaffinity purification procedure for SV40 large T antigen. , 1985, Virology.

[68]  M. Kirschner,et al.  Regulation of the cell cycle during early Xenopus development , 1984, Cell.

[69]  M. Lohka,et al.  Roles of cytosol and cytoplasmic particles in nuclear envelope assembly and sperm pronuclear formation in cell-free preparations from amphibian eggs , 1984, The Journal of cell biology.

[70]  James M. Roberts,et al.  p27Kip1, a cyclin-Cdk inhibitor, links transforming growth factor-beta and contact inhibition to cell cycle arrest. , 1994, Genes & development.

[71]  L. Cox.  DNA replication in cell-free extracts from Xenopus eggs is prevented by disrupting nuclear envelope function. , 1992, Journal of cell science.

[72]  T. Hunt,et al.  Cyclins and their partners during Xenopus oocyte maturation. , 1991, Cold Spring Harbor symposia on quantitative biology.