The human kinesin-like protein RB6K is under tight cell cycle control and is essential for cytokinesis.
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.
Department of Biochemistry, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
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Fontijn
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Goud
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Echard
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Jollivet
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van Marle
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Pannekoek
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Horrevoets
D020121:5' Untranslated Regions; D001483:Base Sequence; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology*; D002455:Cell Division / Q000235:genetics / Q000502:physiology*; D017931:DNA Primers / Q000235:genetics; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D016547:Kinesins / Q000235:genetics* / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008938:Mitosis / Q000235:genetics / Q000502:physiology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation
D020121:5' Untranslated Regions; D017931:DNA Primers; C410089:KIF20A protein, human; C092935:KIF23 protein, human; D008869:Microtubule-Associated Proteins; D012333:RNA, Messenger; D016547:Kinesins