We describe the dynamics of kinetochore dynein-dynactin in living Drosophila embryos and examine the effect of mutant dynein on the metaphase checkpoint. A functional conjugate of dynamitin with green fluorescent protein accumulates rapidly at prometaphase kinetochores, and subsequently migrates off kinetochores towards the poles during late prometaphase and metaphase. This behaviour is seen for several metaphase checkpoint proteins, including Rough deal (Rod). In neuroblasts, hypomorphic dynein mutants accumulate in metaphase and block the normal redistribution of Rod from kinetochores to microtubules. By transporting checkpoint proteins away from correctly attached kinetochores, dynein might contribute to shutting off the metaphase checkpoint, allowing anaphase to ensue.
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ACKNOWLEDGEMENTS We acknowledge helpful discussions with all members of the Hays laboratory, and thank E. D. Salmon for communicating results before publication. This work was supported by grants to T.H. and E.W. from the National Institutes of Health (GM44757 and GM19123 respectively) and the American Heart Association (96002200). R.K., R.B. and F.S. performed this work at the UPR 2420 Centre de Génétique Moléculaire of the CNRS, associated with the Université Pierre et Marie Curie. R.K. was supported in part by grants from the CNRS and from the Association Pour la Recherche sur le Cancer (France). R.B. was supported by grants BD/11488/97 and P/BIA/111055/1998 from FCT, Portugal. F.S. was supported by Le Ministère Nationale de l’Enseignement Supérieur et de la Recherche. Correspondence and requests for materials should be addressed to E.W. Supplementary information is available on Nature Cell Biology’s website (http://cellbio.nature.com).
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