eprintid: 136236 rev_number: 47 eprint_status: archive userid: 608 dir: disk0/00/13/62/36 datestamp: 2010-10-26 01:36:08 lastmod: 2021-10-19 22:23:35 status_changed: 2010-10-26 01:36:08 type: article metadata_visibility: show item_issues_count: 0 creators_name: Takesono, A creators_name: Heasman, SJ creators_name: Wojciak-Stothard, B creators_name: Garg, R creators_name: Ridley, AJ title: Microtubules Regulate Migratory Polarity through Rho/ROCK Signaling in T Cells ispublished: pub divisions: UCL divisions: B02 divisions: C07 keywords: NEUTROPHIL POLARIZATION, ACTIN CYTOSKELETON, ALPHA-TUBULIN, RHO-GTPASES, IN-VIVO, CHEMOTAXIS, MOTILITY, LYMPHOCYTES, DYNAMICS, MYOSIN note: © 2010 Takesono et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. This work was supported by Biotechnology and Biological Sciences Research Council (BBSRC) grant 31/C14420, Leukaemia Research Fund United Kingdom grant 04021 and Cancer Research United Kingdom grant C6620/A8833. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. abstract: Background: 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.Methodology/Principal Findings: 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.Conclusions/Significance: 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 alter T-cell migratory behavior from lamellipodium-based persistent migration to bleb-based migration with frequent turning. date: 2010-01-19 publisher: PUBLIC LIBRARY SCIENCE official_url: http://dx.doi.org/10.1371/journal.pone.0008774 oa_status: green language: eng primo: open primo_central: open_green article_type_text: Article verified: verified_batch elements_source: Web of Science elements_id: 238558 doi: 10.1371/journal.pone.0008774 language_elements: EN lyricists_name: Heasman, Sarah lyricists_id: SHEAS25 full_text_status: public publication: PLOS ONE volume: 5 number: 1 article_number: e8774 issn: 1932-6203 citation: Takesono, A; Heasman, SJ; Wojciak-Stothard, B; Garg, R; Ridley, AJ; (2010) Microtubules Regulate Migratory Polarity through Rho/ROCK Signaling in T Cells. PLOS ONE , 5 (1) , Article e8774. 10.1371/journal.pone.0008774 <https://doi.org/10.1371/journal.pone.0008774>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/136236/1/136236.pdf