RRC ID 30473
Author Baldinger T, Gossen M.
Title Binding of Drosophila ORC proteins to anaphase chromosomes requires cessation of mitotic cyclin-dependent kinase activity.
Journal Mol Cell Biol
Abstract 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.
Volume 29(1)
Pages 140-9
Published 2009-1-1
DOI 10.1128/MCB.00981-08
PII MCB.00981-08
PMID 18955499
PMC PMC2612480
MeSH Anaphase* Animals Animals, Genetically Modified Chromosomes / metabolism* Cyclin-Dependent Kinases / metabolism* Drosophila Proteins / metabolism* Drosophila melanogaster / cytology* Drosophila melanogaster / embryology Drosophila melanogaster / enzymology* Embryo, Nonmammalian / cytology Embryo, Nonmammalian / metabolism Genetic Complementation Test Green Fluorescent Proteins / metabolism Intracellular Space / metabolism Origin Recognition Complex / metabolism* Protein Binding Protein Transport Recombinant Fusion Proteins / metabolism Transgenes
IF 3.611
Times Cited 24
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY CELL BIOLOGY
Resource
Drosophila DGRC#109607 DGRC#109609