Dynamic properties of APC-decorated microtubules in living cells.
The adenomatous polyposis coli (APC) tumour suppressor protein is a component of the Wnt signalling pathway in which it plays a major role in controlling nuclear accumulation of beta-catenin and hence in the modulation of beta-catenin-regulated gene transcription. APC also associates with microtubules at the ends of cytoplasmic extensions in epithelial cells, a distribution that can be reproduced in COS cells ectopically expressing APC. To examine the effect of APC on microtubule properties, we monitored directly the behaviour of APC and of APC-decorated microtubules by time-lapse imaging of cytoplasmic extensions in live COS cells expressing APC tagged with a green fluorescent protein. On the proximal part of microtubules, APC was visualised as particulate material moving unidirectionally towards the plus end of microtubules. The distal parts of microtubules were uniformly decorated by APC and were animated by a motile behaviour in the form of aperiodic bending. This behaviour is likely to be the consequence of compression forces acting on microtubules encountering obstacles while elongating. The majority of APC-decorated microtubules in transfected COS cells was sensitive to depolymerisation by nocodazole, but they contained detyrosinated and acetylated alpha-tubulin, suggesting a reduction in the rate of subunit exchange at their growing end. Taken together, these results demonstrate that microtubule domains uniformly decorated by APC display dynamic and motile properties that may be significant for the postulated role of APC in targeting microtubules to specialised membrane sites.
Dayanandan
Rejith
R
Department of Neuroscience, Institute of Psychiatry, King's College London, United Kingdom.
Butler
Rachel
R
Gordon-Weeks
Phillip R
PR
Matus
Andrew
A
Kaech
Stefanie
S
Lovestone
Simon
S
Anderton
Brian H
BH
Gallo
Jean-Marc
JM
D025601:Adenomatous Polyposis Coli Protein / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D017491:Genes, APC; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D009068:Movement; D011485:Protein Binding; D013997:Time Factors; D014162:Transfection
D025601:Adenomatous Polyposis Coli Protein; D008164:Luminescent Proteins; D008869:Microtubule-Associated Proteins; D049452:Green Fluorescent Proteins