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

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Azoitei, ML; Noh, J; Marston, DJ; Roudot, P; Marshall, CB; Daugird, TA; Lisanza, SL; Sandí, M-J; Ikura, M; Sondek, J; Rottapel, R; Hahn, KM; Danuser, G

Published Date

  • September 2, 2019

Published In

Volume / Issue

  • 218 / 9

Start / End Page

  • 3077 - 3097

PubMed ID

  • 31420453

Pubmed Central ID

  • 31420453

Electronic International Standard Serial Number (EISSN)

  • 1540-8140

Digital Object Identifier (DOI)

  • 10.1083/jcb.201812073

Language

  • eng

Conference Location

  • United States