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Phosphorylation of DNA polymerase α-primase by Cyclin A-dependent kinases regulates initiation of DNA replication in vitro - Oncogene

  • ️Nasheuer, H-P
  • ️Thu Apr 03 1997
  • Original Paper
  • Published: 03 April 1997

Oncogene volume 14, pages 1611–1615 (1997)Cite this article

Abstract

DNA polymerase α-primase is the only known eukaryotic enzyme that can start DNA replication de novo. In this study, we investigated the regulation of DNA replication by phosphorylation of DNA polymerase α-primase. The p180 and the p68 subunits of DNA polymerase α-primase were phosphorylated using Cyclin A-, B- and E- dependent kinases. This phosphorylation did not influence its DNA polymerase activity on activated DNA, but slightly stimulated primase activity using poly(dT) single-stranded DNA (ssDNA) without changing the product length of primers. In contrast, site-specific initiation of replication on plasmid DNA containing the SV40 origin is affected: Cyclin A-Cdk2 and Cyclin A-Cdc2 inhibited initiation of SV40 DNA replication in vitro, Cyclin B-Cdc2 had no effect and Cyclin E-Cdk2 stimulated the initation reaction. DNA polymerase α-primase that was pre-phosphorylated by Cyclin A-Cdk2 was completely unable to initiate the SV40 DNA replication in vitro; Cyclin B-Cdc2-phosphorylated enzyme was moderately inhibited, while Cyclin E-Cdk2-treated DNA polymerase α-primase remained fully active in the initiation reaction.

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Authors and Affiliations

  1. Institut für Biochemie, LMU München, Würmtalstr. 221, München, 81375, Germany

    C Voitenleitner

  2. Department of Molecular Biology, Vanderbilt University, Nashville, 37235, Tennessee, USA

    E Fanning

  3. Abteilung Biochemie, Institut für Molekulare Biotechnologie, Beutenbergerstrasse 11, Jena, 07745, Germany

    H-P Nasheuer

Authors

  1. C Voitenleitner

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  2. E Fanning

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  3. H-P Nasheuer

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Cite this article

Voitenleitner, C., Fanning, E. & Nasheuer, HP. Phosphorylation of DNA polymerase α-primase by Cyclin A-dependent kinases regulates initiation of DNA replication in vitro. Oncogene 14, 1611–1615 (1997). https://doi.org/10.1038/sj.onc.1200975

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  • Received: 15 February 1996

  • Revised: 22 November 1996

  • Accepted: 25 November 1996

  • Issue Date: 03 April 1997

  • DOI: https://doi.org/10.1038/sj.onc.1200975

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