Weingartner, M., Binarova, P., Drykova, D., Schweighofer, A., David, J. P., Heberle-Bors, E., Doonan, J. and Bogre, L. (2001) Dynamic recruitment of Cdc2 to specific microtubule structures during mitosis. Plant Cell, 13 (8).
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A-type cyclin-dependent kinases (CDKs), also known as cdc2, are central to the orderly progression of the cell cycle. We made a functional Green Fluorescent Protein (GFP) fusion with CDK-A (Cdc2-GFP) and followed its subcellular localization during the cell cycle in tobacco cells. During interphase, the Cdc2-GFP fusion protein was found in both the cytoplasm and the nucleus, where it was highly resistant to extraction. In premitotic cells, a bright and narrow equatorial band appeared on the cell surface, resembling the late preprophase band, which disintegrated within 10 min as followed by time-lapse images. Cdc2-GFP was not found on prophase spindles but left the chromatin soon after this stage and associated progressively with the metaphase spindle in a microtubule-dependent manner. Arresting cells in mitosis through the stabilization of microtubules by taxol further enhanced the spindle-localized pool of Cdc2-GFP. Toward the end of mitosis, Cdc2-GFP was found at the midzone of the anaphase spindle and phragmoplast; eventually, it became focused at the midline of these microtubule structures. In detergent-extracted cells, the Cdc2-GFP remained associated with mitotic structures. Retention on spindles was prevented by pretreatment with the CDK-specific inhibitor roscovitine and was enhanced by the protein phosphatase inhibitor okadaic acid. Furthermore, we demonstrate that both the endogenous CDK-A and Cdc2-GFP were cosedimented with taxol-stabilized plant microtubules from cell extracts and that Cdc2 activity was detected together with a fraction of polymerized tubulin. These data provide evidence that the A-type CDKs associate physically with mitotic structures in a microtubule-dependent manner and may be involved in regulating the behavior of specific microtubule arrays throughout mitosis.
This is a Submitted version This version's date is: 2001 This item is not peer reviewed
https://repository.royalholloway.ac.uk/items/62ed69d5-8495-faa8-19c5-978d3ff7534f/3/
Deposited by Research Information System (atira) on 27-Jan-2013 in Royal Holloway Research Online.Last modified on 27-Jan-2013
1040-4651 (Print) 1040-4651 (Linking) Journal Article Research Support, Non-U.S. Gov't