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Cited by 61Protein Kinase Regulation and GTPase SignalingCellular Mechanics and InteractionsMicrotubule and mitosis dynamics

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

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Azoitei, Mihai L., Noh, Jungsik, Marston, Daniel J., Roudot, Philippe, Marshall, Christopher B., Daugird, Timothy A., Lisanza, Sidney L., Sandí, María-José +5 more·2019·Journal of Cell Biology
DOI 10.1083/jcb.201812073PMC PMC6719461
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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.

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  • RhoA activation by microtubule-destabilizing agents depends on GEF-H1.

    “This effect depends on the RhoA activator guanine nucleotide exchange factor H1 (GEF-H1), which is sequestered on microtubules in an inactive form, but becomes active upon release from microtubules 18-22 .”

    From Taxane-induced conformational changes in the microtubule lattice activate GEF-H1-dependent RhoA signalling (2026) · cites this paper as [21]

    The Results section, “Local changes in the MT cytoskeleton regulate GEF-H1 activity” (page unavailable), reports that nocodazole-induced microtubule depolymerization elevated GEF-H1 activity. The Introduction identifies GEF-H1 as a regulator of RhoA activation, but the cited passages do not establish that RhoA activation caused by a microtubule-destabilizing agent depends on GEF-H1. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

  • GEF-H1 can control RhoA activity in response to local changes in the density of dynamic microtubules near the cell margin, e.g., to promote cell invasion or trigger cell retraction and help migrating dendritic cells maintain coherence.

    “In this way, GEF-H1 can control RhoA activity in response to local changes in the density of dynamic microtubules near the cell margin, for example, to promote cell invasion into a dense matrix or trigger cell retraction and help cells with long and elaborate protrusions, such as migrating dendritic cells, to maintain their coherence 21,24-26 .”

    From Taxane-induced conformational changes in the microtubule lattice activate GEF-H1-dependent RhoA signalling (2026) · cites this paper as [21]

    The Discussion (page not provided) says local microtubule disassembly increases GEF-H1 activity, while the Results link GEF-H1 and RhoA activity to cell-edge retraction in migrating MDA-MB-231 cells. Our copy does not establish that this mechanism promotes invasion into a dense matrix or helps migrating dendritic cells maintain coherence. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • GEF-H1 is sequestered on microtubules in an inactive form and becomes active upon release from microtubules.

    “This effect depends on the RhoA activator guanine nucleotide exchange factor H1 (GEF-H1), which is sequestered on microtubules in an inactive form, but becomes active upon release from microtubules 18-22 .”

    From Taxane-induced conformational changes in the microtubule lattice activate GEF-H1-dependent RhoA signalling (2026) · cites this paper as [21]

  • Microtubule-destabilizing agents activate GEF-H1 by inducing its unbinding from microtubules.

    “As microtubule-destabilizing agents activate GEF-H1 by inducing its unbinding from microtubules 18-21”

    From Taxane-induced conformational changes in the microtubule lattice activate GEF-H1-dependent RhoA signalling (2026) · cites this paper as [21]

  • GEFH1 is a critical microtubule-regulated activator of Rho signaling pathways.

    “GEFH1; also known as Arhgef2 or Lfc), a critical microtubule-regulated activator of Rho signaling pathways.”

    From Structural basis of microtubule-mediated signal transduction (2026) · cites this paper as [12]

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Authors
Azoitei, M. L., Noh, J., Marston, D. J., Roudot, P., Marshall, C. B., Daugird, T. A., Lisanza, S. L., Sandí, M.-J., Ikura, M., Sondek, J., Rottapel, R., Hahn, K. M., & Danuser, G.
Affiliations
  • University of North Carolina at Chapel Hill (US)
  • The University of Texas Southwestern Medical Center (US)
  • University Health Network (CA)
  • Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada 3
Corresponding author
  • Hahn, Klaus M.
  • Danuser, Gaudenz
ORCID
Venue
Journal of Cell Biology
Publisher
Rockefeller University Press
Published
2019-08-16
Volume
218
Issue
9
Pages
3077-3097
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
DOI
10.1083/jcb.201812073
PMC
PMC6719461
OpenAlex
W2967825571
Cited by
61
References
56
Citations by year
2026: 9 · 2025: 10 · 2024: 8 · 2023: 12 · 2022: 10 · 2021: 3 · 2020: 8 · 2019: 1
FWCI
1.82
Type
journal article
Language
en
Open access
Licence
Locations
  • The Journal of Cell Biology (publishedVersion, cc-by-nc-sa)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
Funders
National Institutes of Health (R01GM071868, R35GM122596, R01HL133668)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
Keywords
GEF-H1, RhoA activation, Rho GTPases, guanine nucleotide exchange factor, microtubule dynamics, Src kinase phosphorylation, cell migration, cell edge, cell protrusions, cell cortex, actin dynamics, spatiotemporal signaling
MeSH terms
Animals, Chlorocebus aethiops, Humans, Microtubules, Biosensing Techniques, Signal Transduction, src-Family Kinases, COS Cells, rhoA GTP-Binding Protein, HEK293 Cells, Rho Guanine Nucleotide Exchange Factors

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