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MBC in Press, published online ahead of print January 19, 2005
Mol. Biol. Cell 10.1091/mbc.E04-10-0935

A more recent version of this article appeared on April 1, 2005
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Submitted on October 27, 2004
Accepted on January 11, 2005

A Minus-End Directed Kinesin with +TIP Activity Is Involved in Spindle Morphogenesis

J. Christian Ambrose,*{dagger} Wuxing Li,{ddagger} Adam Marcus,{sect}|| Hong Ma,*{dagger}{ddagger}{sect} and Richard Cyr{dagger}{ddagger}{sect}

{sect}Department of Biology, *The Huck Institutes of the Life Sciences, {dagger}Integrative Biosciences Graduate Degree Program, and {ddagger}Plant Physiology Program, The Pennsylvania State University, University Park, PA 16802

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.

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

Corresponding author. E-mail: rjc8{at}psu.edu




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