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APC is a component of an organizing template for cortical microtubule networks

Nature Cell Biology volume 7, pages 463–473 (2005)Cite this article

Abstract

A microtubule network on the basal cortex of polarized epithelial cells consists of non-centrosomal microtubules of mixed polarity. Here, we investigate the proteins that are involved in organizing this network, and we show that end-binding protein 1 (EB1), adenomatous polyposis coli protein (APC) and p150Glued — although considered to be microtubule plus-end-binding proteins — are localized along the entire length of microtubules within the network, and at T-junctions between microtubules. The network shows microtubule behaviours that arise from physical interactions between microtubules, including microtubule plus-end stabilization on the sides of other microtubules, and sliding of microtubule ends along other microtubules. APC also localizes to the basal cortex. Microtubules grew over and paused at APC puncta; an in vitro reconstituted microtubule network overlaid APC puncta; and microtubule network reconstitution was inhibited by function-blocking APC antibodies. Thus, APC is a component of a cortical template that guides microtubule network formation.

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Figure 1: Microtubules interact with one another to cause bending, sliding and pausing.
Figure 2: APC associates both with microtubules and the basal cortex.
Figure 3: APC aligns with microtubules on the basal cortex in cell lines that express APC without the C-terminal microtubule-binding domain.
Figure 4: Distributions of KAP3, p150Glued and microtubules on MDCK and Caco-2 cell basal patches.
Figure 5: Endogenous EB1 binds along the length of microtubules on basal patches of MDCK and Caco-2 cells.
Figure 6: Retrospective staining of APC after imaging microtubule dynamics.
Figure 7: Reconstitution of the microtubule network on isolated membrane patches.
Figure 8: An APC antibody inhibits microtubule polymerization on the basal cortex.

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Acknowledgements

We thank A. Barth for discussions, S. Yamada for discussions and assistance with image processing, P. Coulam for production of affinity-purified APC antibodies, and A. Chhabra for help with data analysis. This work was supported by Postdoctoral Fellowship Grant (PF-03-016-01-CSM) from the American Cancer Society and PHS (5T32CA09151) from the National Cancer Institute, DHHS to A.R., and an NIH grant to W.J.N. (NS 42735).

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  1. Department of Molecular and Cellular Physiology, Beckman Center for Molecular and Genetic Medicine B121, 279 Campus Drive, Stanford University School of Medicine, Stanford, 94305-5435, CA, USA

    Amy Reilein & W. James Nelson

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Correspondence to W. James Nelson.

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Reilein, A., Nelson, W. APC is a component of an organizing template for cortical microtubule networks. Nat Cell Biol 7, 463–473 (2005). https://doi.org/10.1038/ncb1248

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