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Binding of Drosophila ORC proteins to anaphase chromosomes requires cessation of mitotic cyclin-dependent kinase activity

Item Type:Article
Title:Binding of Drosophila ORC proteins to anaphase chromosomes requires cessation of mitotic cyclin-dependent kinase activity
Creators Name:Baldinger, T. and Gossen, M.
Abstract:The initial step in the acquisition of replication competence by eukaryotic chromosomes is the binding of the multi-subunit origin recognition complex, ORC. We describe a transgenic Drosophila model which enables dynamic imaging of a fluorescent protein-tagged DmORC 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 localisation 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 CDK activity is essential for binding of the DmOrc proteins to chromosomes.
Keywords:Anaphase, Genetically Modified Animals, Chromosomes, Cyclin-Dependent Kinases, Drosophila Proteins, Nonmammalian Embryo, Genetic Complementation Test, Green Fluorescent Proteins, Intracellular Space, Origin Recognition Complex, Protein Binding, Protein Transport, Recombinant Fusion Proteins, Transgenes, Animals, Drosophila melanogaster
Source:Molecular and Cellular Biology
ISSN:0270-7306
Publisher:American Society for Microbiology (U.S.A.)
Volume:29
Number:1
Page Range:140-149
Date:January 2009
Official Publication:https://doi.org/10.1128/MCB.00981-08
PubMed:View item in PubMed

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