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Cancers 2016, 8(5), 47; doi:10.3390/cancers8050047

Characterization of Adenomatous Polyposis Coli Protein Dynamics and Localization at the Centrosome

1
Centre for Cancer Research, The Westmead Institute for Medical Research, The University of Sydney, Westmead, NSW 2145, Australia
2
School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong SAR, China
3
Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, China
*
Author to whom correspondence should be addressed.
Academic Editors: Renée van Amerongen and Walter Birchmeier
Received: 14 March 2016 / Revised: 20 April 2016 / Accepted: 26 April 2016 / Published: 30 April 2016
(This article belongs to the Special Issue Wnt Signaling in Cancer)
View Full-Text   |   Download PDF [3043 KB, uploaded 30 April 2016]   |  

Abstract

The adenomatous polyposis coli (APC) tumor suppressor is a multifunctional regulator of Wnt signaling and acts as a mobile scaffold at different cellular sites. APC was recently found to stimulate microtubule (MT) growth at the interphase centrosome; however, little is known about its dynamics and localization at this site. To address this, we analysed APC dynamics in fixed and live cells by fluorescence microscopy. In detergent-extracted cells, we discovered that APC was only weakly retained at the centrosome during interphase suggesting a rapid rate of exchange. This was confirmed in living cells by fluorescence recovery after photobleaching (FRAP), which identified two pools of green fluorescent protein (GFP)-APC: a major rapidly exchanging pool (~86%) and minor retained pool (~14%). The dynamic exchange rate of APC was unaffected by C-terminal truncations implicating a targeting role for the N-terminus. Indeed, we mapped centrosome localization to N-terminal armadillo repeat (ARM) domain amino acids 334–625. Interestingly, the rate of APC movement to the centrosome was stimulated by intact MTs, and APC dynamics slowed when MTs were disrupted by nocodazole treatment or knockdown of γ-tubulin. Thus, the rate of APC recycling at the centrosome is enhanced by MT growth, suggesting a positive feedback to stimulate its role in MT growth. View Full-Text
Keywords: APC; centrosome; colorectal cancer; FRAP; protein dynamics; Wnt signaling APC; centrosome; colorectal cancer; FRAP; protein dynamics; Wnt signaling
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Lui, C.; Mok, M.T.S.; Henderson, B.R. Characterization of Adenomatous Polyposis Coli Protein Dynamics and Localization at the Centrosome. Cancers 2016, 8, 47.

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