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Eukaryotic Cell, December 2007, p. 2354-2364, Vol. 6, No. 12
1535-9778/07/$08.00+0     doi:10.1128/EC.00128-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Kinesin-13 Regulates Flagellar, Interphase, and Mitotic Microtubule Dynamics in Giardia intestinalis{triangledown} ,{dagger}

Scott C. Dawson,1,2* Meredith S. Sagolla,1 Joel J. Mancuso,1 David J. Woessner,2 Susan A. House,2 Lillian Fritz-Laylin,1 and W. Zacheus Cande1

Department of Molecular and Cell Biology, University of California-Berkeley, 341 LSA Bldg., Berkeley, California 94720,1 Section of Microbiology, 255 Briggs Hall, University of California-Davis, Davis, California 956162

Received 17 April 2007/ Accepted 18 July 2007

Microtubule depolymerization dynamics in the spindle are regulated by kinesin-13, a nonprocessive kinesin motor protein that depolymerizes microtubules at the plus and minus ends. Here we show that a single kinesin-13 homolog regulates flagellar length dynamics, as well as other interphase and mitotic dynamics in Giardia intestinalis, a widespread parasitic diplomonad protist. Both green fluorescent protein-tagged kinesin-13 and EB1 (a plus-end tracking protein) localize to the plus ends of mitotic and interphase microtubules, including a novel localization to the eight flagellar tips, cytoplasmic anterior axonemes, and the median body. The ectopic expression of a kinesin-13 (S280N) rigor mutant construct caused significant elongation of the eight flagella with significant decreases in the median body volume and resulted in mitotic defects. Notably, drugs that disrupt normal interphase and mitotic microtubule dynamics also affected flagellar length in Giardia. Our study extends recent work on interphase and mitotic kinesin-13 functioning in metazoans to include a role in regulating flagellar length dynamics. We suggest that kinesin-13 universally regulates both mitotic and interphase microtubule dynamics in diverse microbial eukaryotes and propose that axonemal microtubules are subject to the same regulation of microtubule dynamics as other dynamic microtubule arrays. Finally, the present study represents the first use of a dominant-negative strategy to disrupt normal protein function in Giardia and provides important insights into giardial microtubule dynamics with relevance to the development of antigiardial compounds that target critical functions of kinesins in the giardial life cycle.

* Corresponding author. Mailing address: Section of Microbiology, University of California-Davis, 255 Briggs Hall, One Shields Ave., Davis, CA 95616. Phone: (530) 752-3633. Fax: (530) 752-9014. E-mail: scdawson{at}ucdavis.edu

{triangledown} Published ahead of print on 31 August 2007.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

Eukaryotic Cell, December 2007, p. 2354-2364, Vol. 6, No. 12
1535-9778/07/$08.00+0     doi:10.1128/EC.00128-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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