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Molecular and Cellular Biology, April 2001, p. 2944-2955, Vol. 21, No. 8
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.8.2944-2955.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

The Human Kinesin-Like Protein RB6K Is under Tight Cell Cycle Control and Is Essential for Cytokinesis

Ruud D. Fontijn,1 Bruno Goud,2 Arnaud Echard,2 Florence Jollivet,2 Jan van Marle,3 Hans Pannekoek,1 and Anton J. G. Horrevoets1,*

Departments of Biochemistry1 and Electron Microscopy,3 Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands, and UMR CNRS 144, Institut Curie, Paris, France2

Received 21 September 2000/Returned for modification 25 October 2000/Accepted 10 January 2001

Several members of the kinesin superfamily are known to play a prominent role in the motor-driven transport processes that occur in mitotic cells. Here we describe a new mitotic human kinesin-like protein, RB6K (Rabkinesin 6), distantly related to MKLP-1. Expression of RB6K is regulated during the cell cycle at both the mRNA and protein level and, similar to cyclin B, shows a maximum during M phase. Isolation of the RB6K promoter allowed identification of a CDE-CHR element and promoter activity was shown to be maximal during M phase. Immunofluorescence microscopy using antibodies raised against RB6K showed a weak signal in interphase Golgi but a 10-fold higher signal in prophase nuclei. During M phase, the newly synthesized RB6K does not colocalise with Rab6. In later stages of mitosis RB6K localized to the spindle midzone and appeared on the midbodies during cytokinesis. The functional significance of this localization during M phase was revealed by antibody microinjection studies which resulted exclusively in binucleate cells, showing a complete failure of cytokinesis. These results substantiate a crucial role for RB6K in late anaphase B and/or cytokinesis, clearly distinct from the role of MKLP-1.

* Corresponding author. Mailing address: Department of Biochemistry, Academic Medical Center K1-160, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. Phone: 31-20-5665124. Fax: 31-20-6915519. E-mail: a.j.horrevoets{at}amc.uva.nl.

Molecular and Cellular Biology, April 2001, p. 2944-2955, Vol. 21, No. 8
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.8.2944-2955.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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