Please use this identifier to cite or link to this item: https://doi.org/10.1242/jcs.110494
Title: Microtubule guidance tested through controlled cell geometry
Authors: Huda, S
Soh, S 
Pilans, D
Byrska-Bishop, M
Kim, J
Wilk, G
Borisy, G.G
Kandere-Grzybowska, K
Grzybowski, B.A
Keywords: F actin
fibronectin
green fluorescent protein
myosin IIA
myosin IIB
animal cell
article
cell function
cell growth
cell nucleus
cell shape
cell structure
centrosome
controlled study
female
focal adhesion
gene deletion
human
human cell
microtubule
microtubule guidance
nonhuman
priority journal
quantitative analysis
rat
RNA interference
Actins
Animals
Cell Line
HeLa Cells
Humans
Microtubules
Myosin Type II
Rats
RNA Interference
Issue Date: 2012
Citation: Huda, S, Soh, S, Pilans, D, Byrska-Bishop, M, Kim, J, Wilk, G, Borisy, G.G, Kandere-Grzybowska, K, Grzybowski, B.A (2012). Microtubule guidance tested through controlled cell geometry. Journal of Cell Science 125 (23) : 5790-5799. ScholarBank@NUS Repository. https://doi.org/10.1242/jcs.110494
Rights: Attribution 4.0 International
Abstract: In moving cells dynamic microtubules (MTs) target and disassemble substrate adhesion sites (focal adhesions; FAs) in a process that enables the cell to detach from the substrate and propel itself forward. The short-range interactions between FAs and MT plus ends have been observed in several experimental systems, but the spatial overlap of these structures within the cell has precluded analysis of the putative long-range mechanisms by which MTs growing through the cell body reach FAs in the periphery of the cell. In the work described here cell geometry was controlled to remove the spatial overlap of cellular structures thus allowing for unambiguous observation of MT guidance. Specifically, micropatterning of living cells was combined with high-resolution in-cell imaging and gene product depletion by means of RNA interference to study the long-range MT guidance in quantitative detail. Cells were confined on adhesive triangular microislands that determined cell shape and ensured that FAs localized exclusively at the vertices of the triangular cells. It is shown that initial MT nucleation at the centrosome is random in direction, while the alignment of MT trajectories with the targets (i.e. FAs at vertices) increases with an increasing distance from the centrosome, indicating that MT growth is a non-random, guided process. The guided MT growth is dependent on the presence of FAs at the vertices. The depletion of either myosin IIA or myosin IIB results in depletion of F-actin bundles and spatially unguided MT growth. Taken together our findings provide quantitative evidence of a role for long-range MT guidance in MT targeting of FAs. © 2012. Published by The Company of Biologists Ltd.
Source Title: Journal of Cell Science
URI: https://scholarbank.nus.edu.sg/handle/10635/180813
ISSN: 0021-9533
DOI: 10.1242/jcs.110494
Rights: Attribution 4.0 International
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