Spatiotemporal dynamics of GEF-H1 activation controlled by microtubule- and Src-mediated pathways

Mihai L. Azoitei, Jungsik Noh, Daniel J. Marston, Philippe Roudot, Christopher B. Marshall, Timothy A. Daugird, Sidney L. Lisanza, María José Sandí, Mitsu Ikura, John Sondek, Robert Rottapel, Klaus M. Hahn, Gaudenz Danuser

Research output: Contribution to journalArticle

Abstract

Rho family GTPases are activated with precise spatiotemporal control by guanine nucleotide exchange factors (GEFs). Guanine exchange factor H1 (GEF-H1), a RhoA activator, is thought to act as an integrator of microtubule (MT) and actin dynamics in diverse cell functions. Here we identify a GEF-H1 autoinhibitory sequence and exploit it to produce an activation biosensor to quantitatively probe the relationship between GEF-H1 conformational change, RhoA activity, and edge motion in migrating cells with micrometer- and second-scale resolution. Simultaneous imaging of MT dynamics and GEF-H1 activity revealed that autoinhibited GEF-H1 is localized to MTs, while MT depolymerization subadjacent to the cell cortex promotes GEF-H1 activation in an ~5-µm-wide peripheral band. GEF-H1 is further regulated by Src phosphorylation, activating GEF-H1 in a narrower band ~0-2 µm from the cell edge, in coordination with cell protrusions. This indicates a synergistic intersection between MT dynamics and Src signaling in RhoA activation through GEF-H1.

Original languageEnglish (US)
Pages (from-to)3077-3097
Number of pages21
JournalThe Journal of cell biology
Volume218
Issue number9
DOIs
StatePublished - Sep 2 2019

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Guanine Nucleotide Exchange Factors
Microtubules
rho GTP-Binding Proteins
Guanine
Biosensing Techniques
Actins
Phosphorylation

ASJC Scopus subject areas

  • Cell Biology

Cite this

Spatiotemporal dynamics of GEF-H1 activation controlled by microtubule- and Src-mediated pathways. / Azoitei, Mihai L.; Noh, Jungsik; Marston, Daniel J.; Roudot, Philippe; Marshall, Christopher B.; Daugird, Timothy A.; Lisanza, Sidney L.; Sandí, María José; Ikura, Mitsu; Sondek, John; Rottapel, Robert; Hahn, Klaus M.; Danuser, Gaudenz.

In: The Journal of cell biology, Vol. 218, No. 9, 02.09.2019, p. 3077-3097.

Research output: Contribution to journalArticle

Azoitei, ML, Noh, J, Marston, DJ, Roudot, P, Marshall, CB, Daugird, TA, Lisanza, SL, Sandí, MJ, Ikura, M, Sondek, J, Rottapel, R, Hahn, KM & Danuser, G 2019, 'Spatiotemporal dynamics of GEF-H1 activation controlled by microtubule- and Src-mediated pathways', The Journal of cell biology, vol. 218, no. 9, pp. 3077-3097. https://doi.org/10.1083/jcb.201812073
Azoitei, Mihai L. ; Noh, Jungsik ; Marston, Daniel J. ; Roudot, Philippe ; Marshall, Christopher B. ; Daugird, Timothy A. ; Lisanza, Sidney L. ; Sandí, María José ; Ikura, Mitsu ; Sondek, John ; Rottapel, Robert ; Hahn, Klaus M. ; Danuser, Gaudenz. / Spatiotemporal dynamics of GEF-H1 activation controlled by microtubule- and Src-mediated pathways. In: The Journal of cell biology. 2019 ; Vol. 218, No. 9. pp. 3077-3097.
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