RNAi of mitotic cyclins in Drosophila uncouples the nuclear and centrosome cycle.
BACKGROUND
Successful cell duplication requires orderly progression through a succession of dramatic cell-cycle events. Disruption of this precise coupling can compromise genomic integrity. The coordination of cell-cycle events is thought to arise from control by a single master regulator, cyclin:Cdk, whose activity oscillates. However, we still know very little of how individual cell-cycle events are coupled to this oscillator and how the timing of each event is controlled.


RESULTS
We developed an approach with RNA interference (RNAi) and real-time imaging to study cyclin contributions to the rapid syncytial divisions of Drosophila embryos. Simultaneous knockdown of all three mitotic cyclins blocked nuclei from entering mitosis. Despite nuclear arrest, centrosomes and associated myosin cages continued to divide until the midblastula transition. Centrosome division was synchronous throughout the embryo and the period of the uncoupled duplication cycle increased over successive divisions. In contrast to its normal actions, injection of a competitive inhibitor of the anaphase-promoting complex/cyclosome (APC/C) after knockdown of the mitotic cyclins did not interfere with the centrosome-duplication cycles. Finally, we examined how cyclin knockdown affects the onset of cellularization at the midblastula transition and found that nuclear cell-cycle arrest did not advance or delay onset of cellularization.


CONCLUSIONS
We show that knockdown of mitotic cyclins allows centrosomes to duplicate in a cycle that is uncoupled from other cell-cycle events. We suggest that high mitotic cyclin normally ensures that the centrosome cycle remains entrained to the nuclear cycle.
McCleland
Mark L
ML
Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California 94143-2200, USA.
O'Farrell
Patrick H
PH
D000818:Animals; D001756:Blastoderm / Q000378:metabolism*; D018385:Centrosome / Q000502:physiology*; D003078:Colchicine; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D016213:Cyclins / Q000378:metabolism*; D003513:Cycloheximide; D004330:Drosophila / Q000196:embryology* / Q000378:metabolism; D008938:Mitosis / Q000502:physiology*; D009218:Myosins / Q000378:metabolism; D011500:Protein Synthesis Inhibitors; D034622:RNA Interference; D050257:Tubulin Modulators
D016213:Cyclins; D011500:Protein Synthesis Inhibitors; D050257:Tubulin Modulators; D003513:Cycloheximide; D018844:Cyclin-Dependent Kinases; D009218:Myosins; D003078:Colchicine