Binding of Drosophila Orc Proteins to Anaphase Chromosomes Requires Cessation of Mitotic Cyclin-Dependent Kinase Activity
| Autor: | Tina Baldinger, Manfred Gossen |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Embryo
Nonmammalian Recombinant Fusion Proteins Green Fluorescent Proteins Cyclin A Intracellular Space Origin Recognition Complex Biology Chromosomes Animals Genetically Modified Animals Drosophila Proteins Transgenes Kinase activity Molecular Biology Mitosis Metaphase Anaphase Genetic Complementation Test fungi Articles Cell Biology Cyclin-Dependent Kinases Chromatin Cell biology Protein Transport Drosophila melanogaster biology.protein Origin recognition complex Drosophila Protein Protein Binding |
| Zdroj: | Molecular and Cellular Biology. 29:140-149 |
| ISSN: | 1098-5549 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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