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MBC in Press, published online ahead of print June 27, 2003
Mol. Biol. Cell 10.1091/mbc.E03-04-0214

A more recent version of this article appeared on October 1, 2003
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Submitted on April 8, 2003
Revised on May 28, 2003
Accepted on May 28, 2003

Localisation of Pavarotti-KLP in living Drosophila embryos suggests roles in re-organising the cortical cytoskeleton during the mitotic cycle

Gianluca Minestrini1, Alyssa S. Harley1, and David M. Glover1*

1 Cancer Research UK, Cell Cycle Genetics Group, University of Cambridge, Department of Genetics, Downing Street, Cambridge CB2 3EH

* Corresponding author. E-mail address: dmg25{at}mole.bio.cam.ac.uk.

Pav-KLP is the Drosophila member of the MKLP1 family essential for cytokinesis. In the syncytial blastoderm embryo GFP-Pav-KLP cyclically associates with astral, spindle and midzone microtubules, and also to actomyosin pseudocleavage furrows. As the embryo cellularises, GFP-Pav-KLP also localizes to the leading edge of the furrows that form cells. In mononucleate cells, nuclear localization of GFP-Pav-KLP is mediated through NLS elements in its C-terminal domain. Mutants in these elements that delocalize Pav-KLP to the cytoplasm in interphase do not affect cell division. In mitotic cells, one population of wild-type GFP-Pav-KLP associates with the spindle and concentrates in the midzone at anaphase B. A second is at the cell cortex on mitotic entry, and later concentrates in the region of the cleavage furrow. An ATP binding mutant does not localize to the cortex and spindle midzone but accumulates on spindle pole microtubules to which actin is recruited. This leads either to failure of the cleavage furrow to form or later defects in which daughter cells remain connected by a microtubule bridge. Together this suggests Pav-KLP transports elements of the actomyosin cytoskeleton to plus ends of astral microtubules in the equatorial region of the cell to permit cleavage ring formation.




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