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Related Experiment Videos

DNA replication and the cell cycle.

B Stillman1, S P Bell, A Dutta

  • 1Cold Spring Harbor Laboratory, New York 11724.

Ciba Foundation Symposium
|January 1, 1992
PubMed
Summary
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Cell cycle regulation of DNA replication involves cdc2 kinase phosphorylating Replication Protein A (RPA). A novel origin recognition complex in yeast binds cellular DNA origins, aiding replication.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Eukaryotic DNA replication is tightly regulated during the cell cycle.
  • Studies on simian virus 40 (SV40) DNA replication identified key human replication proteins.
  • Replication Protein A (RPA) phosphorylation is cell cycle-dependent, starting at DNA replication onset.

Purpose of the Study:

  • To investigate the role of cdc2 protein kinase in regulating DNA replication.
  • To identify and characterize proteins involved in cellular DNA origin binding in yeast.
  • To understand the mechanism of DNA replication initiation.

Main Methods:

  • In vitro phosphorylation assays using cdc2 protein kinase and RPA.
  • Biochemical purification of a multiprotein complex from Saccharomyces cerevisiae.

Related Experiment Videos

  • Analysis of yeast DNA replication origins and protein binding.

    • Main Results:

    • RPA phosphorylation by cdc2 kinase occurs at the start of DNA replication.
    • cdc2 kinase stimulates SV40 DNA origin unwinding in vitro.
    • A multiprotein complex, potentially an origin recognition complex, was identified and purified from yeast.
    • This complex binds to all characterized S. cerevisiae cellular origins.

    Conclusions:

    • cdc2 kinase plays a role in regulating DNA replication initiation.
    • A conserved mechanism for origin recognition exists between viruses and yeast.
    • The identified yeast complex may function as a cellular origin recognition complex, crucial for DNA replication.