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Vol. 17, Issue 5, 2331-2345, May 2006
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* Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037;
Division of Cell and Developmental Biology, School of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom
Submitted June 7, 2005;
Revised January 31, 2006;
Accepted February 24, 2006
Monitoring Editor: Ted Salmon
In interphase cells, the adenomatous polyposis coli (APC) protein accumulates on a small subset of microtubules (MTs) in cell protrusions, suggesting that APC may regulate the dynamics of these MTs. We comicroinjected a nonperturbing fluorescently labeled monoclonal antibody and labeled tubulin to simultaneously visualize dynamics of endogenous APC and MTs in living cells. MTs decorated with APC spent more time growing and had a decreased catastrophe frequency compared with non-APC-decorated MTs. Endogenous APC associated briefly with shortening MTs. To determine the relationship between APC and its binding partner EB1, we monitored EB1-green fluorescent protein and endogenous APC concomitantly in living cells. Only a small fraction of EB1 colocalized with APC at any one time. APC-deficient cells and EB1 small interfering RNA showed that EB1 and APC localized at MT ends independently. Depletion of EB1 did not change the growth-stabilizing effects of APC on MT plus ends. In addition, APC remained bound to MTs stabilized with low nocodazole, whereas EB1 did not. Thus, we demonstrate that the association of endogenous APC with MT ends correlates directly with their increased growth stability, that this can occur independently of its association with EB1, and that APC and EB1 can associate with MT plus ends by distinct mechanisms.
Abbreviations used: APC, adenomatous polyposis coli; MT, microtubule.
The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).
Present address: Department of Cell and Tissue Biology, University of California at San Francisco School of Dentistry, 513 Parnassus Ave., HSW-616, Box 0512, San Francisco, CA 94143-0512.
Address correspondence to: Clare M. Waterman-Storer (waterman{at}scripps.edu).
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