Requirement of Cyclin/Cdk2 and Protein Phosphatase 1 Activity for Chromatin Assembly Factor 1-dependent Chromatin Assembly during DNA Synthesis*

The influence of reversible protein phosphorylation on nucleosome assembly during DNA replication was analyzed in extracts from human cells. Inhibitor studies and add-back experiments indicated requirements of cyclin A/Cdk2, cyclin E/Cdk2, and protein phosphatase type 1 (PP1) activities for nucleosome assembly during DNA synthesis by chromatin assembly factor 1 (CAF-1). The p60 subunit of CAF-1 is a molecular target for reversible phosphorylation by cyclin/Cdk complexes and PP1 during nucleosome assembly and DNA synthesis in vitro. Purified p60 can be directly phosphorylated by purified cyclin A/Cdk2, cyclin E/Cdk2, and cyclin B1/Cdk1, but not by cyclin D/Cdk4 complexes in vitro. Cyclin B1/Cdk1 triggers hyperphosphorylation of p60 in the presence of additional cytosolic factors. CAF-1 containing hyperphosphorylated p60 prepared from mitotic cells is inactive in nucleosome assembly and becomes activated by dephosphorylation in vitro. These data provide functional evidence for a requirement of the cell cycle machinery for nucleosome assembly by CAF-1 during DNA replication.

[1]  J. Pines,et al.  Cyclin/Cdk-Dependent Initiation of DNA Replication in a Human Cell-Free System , 1997, Cell.

[2]  R. Knippers,et al.  Comparison of replicative and non-replicative chromatin assembly pathways in HeLa cell extracts. , 1992, Biochimica et biophysica acta.

[3]  G. Hannon,et al.  Differential effects by the p21 CDK inhibitor on PCNA-dependent DNA replication and repair , 1994, Nature.

[4]  P. Cohen.  The structure and regulation of protein phosphatases. , 1989, Annual review of biochemistry.

[5]  B. Stillman,et al.  Stepwise assembly of chromatin during DNA replication in vitro. , 1991, The EMBO journal.

[6]  A. Philpott,et al.  Regular ArticleNuclear chaperones , 2000 .

[7]  Bruce Stillman,et al.  Nucleosomal DNA regulates the core-histone-binding subunit of the human Hat1 acetyltransferase , 1998, Current Biology.

[8]  J. Li,et al.  Simian virus 40 DNA replication in vitro. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[9]  J. Pines,et al.  Cyclins and cyclin-dependent kinases: a biochemical view. , 1995, The Biochemical journal.

[10]  E. Fanning.  Control of SV40 DNA replication by protein phosphorylation: a model for cellular DNA replication? , 1994, Trends in cell biology.

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

[12]  J. Roberts,et al.  Evolving Ideas about Cyclins , 1999, Cell.

[13]  L. J. Peck,et al.  Energetics of B-to-Z transition in DNA. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[14]  C. Gauss,et al.  Inhibition of the Ser-Thr phosphatases PP1 and PP2A by naturally occurring toxins. , 1997, Bioorganic & medicinal chemistry.

[15]  L Meijer,et al.  Biochemical and cellular effects of roscovitine, a potent and selective inhibitor of the cyclin-dependent kinases cdc2, cdk2 and cdk5. , 1997, European journal of biochemistry.

[16]  J. Pines,et al.  Human Cyclin a Is Required for Mitosis until Mid Prophase , 1999, The Journal of cell biology.

[17]  T. Krude,et al.  Minichromosome replication in vitro: inhibition of re-replication by replicatively assembled nucleosomes. , 1994, The Journal of biological chemistry.

[18]  T. Krude,et al.  Nucleosome assembly during complementary DNA strand synthesis in extracts from mammalian cells. , 1993, The Journal of biological chemistry.

[19]  Bruce Stillman,et al.  The p150 and p60 subunits of chromatin assemblyfactor I: A molecular link between newly synthesized histories and DNA replication , 1995, Cell.

[20]  B. Stillman,et al.  Postreplicative chromatin assembly by Drosophila and human chromatin assembly factor 1 , 1996, Molecular and cellular biology.

[21]  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.

[22]  T. Hunter,et al.  Human cyclins A and B1 are differentially located in the cell and undergo cell cycle-dependent nuclear transport , 1991, The Journal of cell biology.

[23]  B. Stillman,et al.  Chromatin Assembly Coupled to DNA Repair: A New Role for Chromatin Assembly Factor I , 1996, Cell.

[24]  K. Marheineke,et al.  Nucleosome Assembly Activity and Intracellular Localization of Human CAF-1 Changes during the Cell Division Cycle* , 1998, The Journal of Biological Chemistry.

[25]  E. Villa-Moruzzi,et al.  Differential Subcellular Localization of Protein Phosphatase-1 α, γ1, and δ Isoforms during Both Interphase and Mitosis in Mammalian Cells , 1998, The Journal of cell biology.

[26]  T. Krude.  Chromatin assembly during DNA replication in somatic cells. , 1999, European journal of biochemistry.

[27]  P. Chambon,et al.  Folding of the DNA double helix in chromatin-like structures from simian virus 40. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[28]  R. Kamakaka,et al.  Chromatin assembly: biochemical identities and genetic redundancy. , 1999, Current opinion in genetics & development.

[29]  S. Elledge,et al.  Inhibition of cyclin-dependent kinases by p21. , 1995, Molecular biology of the cell.

[30]  J. Blow,et al.  Inhibition of cyclin-dependent kinases by purine analogues. , 1994, European journal of biochemistry.

[31]  C. MacKintosh,et al.  Inhibitors of protein kinases and phosphatases. , 1994, Trends in biochemical sciences.

[32]  D. Lane,et al.  A small peptide inhibitor of DNA replication defines the site of interaction between the cyclin-dependent kinase inhibitor p21WAF1 and proliferating cell nuclear antigen , 1995, Current Biology.

[33]  David O. Morgan,et al.  Principles of CDK regulation , 1995, Nature.

[34]  Z. Kelman,et al.  Protein-PCNA interactions: a DNA-scanning mechanism? , 1998, TIBS -Trends in Biochemical Sciences. Regular ed.

[35]  B. Stillman,et al.  Cell Cycle Control of DNA Replication , 1996, Science.

[36]  T. Krude.  Chromatin assembly factor 1 (CAF-1) colocalizes with replication foci in HeLa cell nuclei. , 1995, Experimental cell research.

[37]  A. Verreault.  De novo nucleosome assembly: new pieces in an old puzzle. , 2000, Genes & development.

[38]  R. Kobayashi,et al.  Nucleosome Assembly by a Complex of CAF-1 and Acetylated Histones H3/H4 , 1996, Cell.

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

[40]  S H Kim,et al.  Inhibition of cyclin-dependent kinases by purine analogues: crystal structure of human cdk2 complexed with roscovitine. , 1997, European journal of biochemistry.

[41]  T. Richmond,et al.  Crystal structure of the nucleosome core particle at 2.8 Å resolution , 1997, Nature.

[42]  E. Lees.  Cyclin dependent kinase regulation. , 1995, Current opinion in cell biology.

[43]  P. Grandi,et al.  A CAF-1–PCNA-Mediated Chromatin Assembly Pathway Triggered by Sensing DNA Damage , 2000, Molecular and Cellular Biology.

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

[45]  D. Brautigan,et al.  Protein phosphatase type 1 in mammalian cell mitosis: chromosomal localization and involvement in mitotic exit , 1992, The Journal of cell biology.

[46]  J. R. Paulson,et al.  Evidence that the endogenous histone H1 phosphatase in HeLa mitotic chromosomes is protein phosphatase 1, not protein phosphatase 2A. , 1996, Journal of cell science.

[47]  B. Ducommun,et al.  Identification of binding domains on the p21Cip1 cyclin-dependent kinase inhibitor. , 1995, Oncogene.

[48]  J. Massagué,et al.  Cell-cycle inhibition by independent CDK and PCNA binding domains in p21Cip1 , 1995, Nature.

[49]  M. Osley.  The regulation of histone synthesis in the cell cycle. , 1991, Annual review of biochemistry.

[50]  T. Krude.  Chromatin replication: Finding the right connection , 1999, Current Biology.

[51]  James M. Roberts,et al.  Inhibitors of mammalian G1 cyclin-dependent kinases. , 1995, Genes & development.

[52]  B. Stillman,et al.  Purification and characterization of CAF-I, a human cell factor required for chromatin assembly during DNA replication in vitro , 1989, Cell.

[53]  E. Nigg,et al.  Cyclin‐dependent protein kinases: Key regulators of the eukaryotic cell cycle , 1995, BioEssays : news and reviews in molecular, cellular and developmental biology.

[54]  B. Stillman,et al.  The N-terminal domains of histones H3 and H4 are not necessary for chromatin assembly factor-1- mediated nucleosome assembly onto replicated DNA in vitro. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[55]  Z. Jónsson,et al.  Proliferating cell nuclear antigen: More than a clamp for DNA polymerases , 1997, BioEssays : news and reviews in molecular, cellular and developmental biology.

[56]  M. Kastan.  Checkpoint controls and cancer , 1997 .

[57]  B. Stillman,et al.  The DNA replication fork in eukaryotic cells. , 1998, Annual review of biochemistry.

[58]  G. Almouzni,et al.  Recruitment of Phosphorylated Chromatin Assembly Factor 1 to Chromatin after UV Irradiation of Human Cells , 1998, The Journal of cell biology.

[59]  T. Krude.  Initiation of Human DNA Replication in Vitro Using Nuclei from Cells Arrested at an Initiation-competent State* , 2000, The Journal of Biological Chemistry.

[60]  M. DePamphilis.  Concepts in eukaryotic DNA replication , 1999 .

[61]  B. Stillman,et al.  Replication-Dependent Marking of DNA by PCNA Facilitates CAF-1-Coupled Inheritance of Chromatin , 1999, Cell.