Microtubules regulate migratory polarity through Rho/ROCK signaling in T cells.

PloS One
Aya TakesonoAnne J Ridley

Abstract

Migrating leukocytes normally have a polarized morphology with an actin-rich lamellipodium at the front and a uropod at the rear. Microtubules (MTs) are required for persistent migration and chemotaxis, but how they affect cell polarity is not known. Here we report that T cells treated with nocodazole to disrupt MTs are unable to form a stable uropod or lamellipodium, and instead often move by membrane blebbing with reduced migratory persistence. However, uropod-localized receptors and ezrin/radixin/moesin proteins still cluster in nocodazole-treated cells, indicating that MTs are required specifically for uropod stability. Nocodazole stimulates RhoA activity, and inhibition of the RhoA target ROCK allows nocodazole-treated cells to re-establish lamellipodia and uropods and persistent migratory polarity. ROCK inhibition decreases nocodazole-induced membrane blebbing and stabilizes MTs. The myosin inhibitor blebbistatin also stabilizes MTs, indicating that RhoA/ROCK act through myosin II to destabilize MTs. Our results indicate that RhoA/ROCK signaling normally contributes to migration by affecting both actomyosin contractility and MT stability. We propose that regulation of MT stability and RhoA/ROCK activity is a mechanism to ...Continue Reading

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May 23, 2014·Cellular and Molecular Life Sciences : CMLS·Yulia ArtemenkoPeter N Devreotes
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Methods Mentioned

BETA
GTPases
pulldown
acetylation
Assay
FCS
FACS
confocal microscopy

Software Mentioned

Bio
Cell Quest
Rad
Mathematica
Kinetic Imaging
ImagePro
LaserPix
LaserSharp

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