Protein-Induced Alterations in DNA Structure at the dhfr Origin of Replication

  • P. Held
  • E. Soultanakis
  • L. Dailey
  • T. Kouzarides
  • N. Heintz
  • N. H. Heintz
Conference paper
Part of the Colloquium der Gesellschaft für Biologische Chemie book series (MOSBACH, volume 43)

Abstract

To identify the trans-acting factors that participate in initiation of DNA synthesis in mammalian cells, we have studied the interaction of nuclear proteins with the DNA sequences that encompass the origin of replication associated with the Chinese hamster dihydrofolate reductase (dhfr) gene. A variety of experimental approaches suggests that replication of amplified dhfr genes in die methotrexate-resistant variant of CHO cells, CHOC 400, initiates within a 4.3 kb Xba I fragment located 14 kb 3’ to the dhfr gene (Heintz and Hamlin 1982; Burhans et al. 1986a, b; Anachakova and Hamlin 1988; Leu and Hamlin 1989). By mapping the strand specificity of Okazaki fragment synthesis throughout the dhfr origin region, an origin of bidirectional DNA synthesis (OBR) has been located to a 450 bp segment of the 4.3 kb Xba I fragment (Burhans et al. 1990).

Keywords

Electrophoresis Polypeptide Polyacrylamide Posite Schiff 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1993

Authors and Affiliations

  • P. Held
    • 1
  • E. Soultanakis
    • 1
  • L. Dailey
    • 2
  • T. Kouzarides
    • 3
  • N. Heintz
    • 2
  • N. H. Heintz
    • 1
  1. 1.Department of Pathology, College of MedicineUniversity of VermontBurlingtonUSA
  2. 2.Laboratory of Molecular BiologyRockefeller UniversityNew YorkUSA
  3. 3.Wellcome/CRC InstituteCambridgeUK

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