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Vol. 16, Issue 4, 1584-1592, April 2005

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A Minus-End–directed Kinesin with Plus-End Tracking Protein Activity Is Involved in Spindle Morphogenesis

J. Christian Ambrose ^* , Wuxing Li , Adam Marcus  ^||, Hong Ma ^*   , and Richard Cyr   

Department of Biology, The Pennsylvania State University, University Park, PA 16802; ^* The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802; Integrative Biosciences Graduate Degree Program, The Pennsylvania State University, University Park, PA 16802; and Plant Physiology Program, The Pennsylvania State University, University Park, PA 16802

Submitted October 27, 2004; Accepted January 11, 2005
Monitoring Editor: J. Richard McIntosh

Diverse kinesin motor proteins are involved in spindle function; however, the mechanisms by which they are targeted to specific sites within spindles are not well understood. Here, we show that a fusion between yellow fluorescent protein (YFP) and a minus-end–directed Kinesin-14 (C-terminal family) from Arabidopsis, ATK5, localizes to mitotic spindle midzones and regions rich in growing plus-ends within phragmoplasts. Notably, in Arabidopsis interphase cells, YFP::ATK5 localizes to microtubules with a preferential enrichment at growing plus-ends; indicating ATK5 is a plus-end tracking protein (+TIP). This +TIP activity is conferred by regions outside of the C-terminal motor domain, which reveals the presence of independent plus-end tracking and minus-end motor activities within ATK5. Furthermore, mitotic spindles of atk5 null mutant plants are abnormally broadened. Based on these data, we propose a model in which ATK5 uses plus-end tracking to reach spindle midzones, where it then organizes microtubules via minus-end–directed motor activity.

Abbreviations used: ATK5, Arabidopsis thaliana kinesin 5; ATK1, Arabidopsis thaliana kinesin 1; YFP, yellow fluorescent protein.

The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

^|| Present address: Emory University, Winship Cancer Institute, 1365C Clifton Rd., Room C4054, Atlanta, GA 30322.

Address correspondence to: Richard Cyr (rjc8{at}psu.edu).

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