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Molecular and Cellular Biology, January 2009, p. 140-149, Vol. 29, No. 1
0270-7306/09/$08.00+0     doi:10.1128/MCB.00981-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Binding of Drosophila Orc Proteins to Anaphase Chromosomes Requires Cessation of Mitotic Cyclin-Dependent Kinase Activity {triangledown} ,{dagger}

Tina Baldinger and Manfred Gossen*

Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany

Received 22 June 2008/ Returned for modification 25 July 2008/ Accepted 20 October 2008

The initial step in the acquisition of replication competence by eukaryotic chromosomes is the binding of the multisubunit origin recognition complex, ORC. We describe a transgenic Drosophila model which enables dynamic imaging of a green fluorescent protein (GFP)-tagged Drosophila melanogaster ORC subunit, DmOrc2-GFP. It is functional in genetic complementation, expressed at physiological levels, and participates quantitatively in complex formation. This fusion protein is therefore able to depict both the holocomplex DmOrc1-6 and the core complex DmOrc2-6 formed by the Drosophila initiator proteins. Its localization can be monitored in vivo along the cell cycle and development. DmOrc2-GFP is not detected on metaphase chromosomes but binds rapidly to anaphase chromatin in Drosophila embryos. Expression of either stable cyclin A, B, or B3 prevents this reassociation, suggesting that cessation of mitotic cyclin-dependent kinase activity is essential for binding of the DmOrc proteins to chromosomes.

* Corresponding author. Mailing address: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Strasse 10, 13125 Berlin, Germany. Phone: 49 (0) 30-9406-2596. Fax: 49 (0) 30-9406-2551. E-mail: mgossen{at}mdc-berlin.de

{triangledown} Published ahead of print on 27 October 2008.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, January 2009, p. 140-149, Vol. 29, No. 1
0270-7306/09/$08.00+0     doi:10.1128/MCB.00981-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Balasov, M., Huijbregts, R. P. H., Chesnokov, I. (2009). Functional analysis of an Orc6 mutant in Drosophila. Proc. Natl. Acad. Sci. USA 106: 10672-10677 [Abstract] [Full Text]  
  • Zhu, W., DePamphilis, M. L. (2009). Selective Killing of Cancer Cells by Suppression of Geminin Activity. Cancer Res. 69: 4870-4877 [Abstract] [Full Text]  


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