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  • Optogenetic Tuning Reveals Rho Amplification-Dependent Dynamics of a Cell Contraction Signal Network.

Optogenetic Tuning Reveals Rho Amplification-Dependent Dynamics of a Cell Contraction Signal Network.

Cell reports (2020-12-03)
Dominic Kamps, Johannes Koch, Victor O Juma, Eduard Campillo-Funollet, Melanie Graessl, Soumya Banerjee, Tomáš Mazel, Xi Chen, Yao-Wen Wu, Stephanie Portet, Anotida Madzvamuse, Perihan Nalbant, Leif Dehmelt
ABSTRACT

Local cell contraction pulses play important roles in tissue and cell morphogenesis. Here, we improve a chemo-optogenetic approach and apply it to investigate the signal network that generates these pulses. We use these measurements to derive and parameterize a system of ordinary differential equations describing temporal signal network dynamics. Bifurcation analysis and numerical simulations predict a strong dependence of oscillatory system dynamics on the concentration of GEF-H1, an Lbc-type RhoGEF, which mediates the positive feedback amplification of Rho activity. This prediction is confirmed experimentally via optogenetic tuning of the effective GEF-H1 concentration in individual living cells. Numerical simulations show that pulse amplitude is most sensitive to external inputs into the myosin component at low GEF-H1 concentrations and that the spatial pulse width is dependent on GEF-H1 diffusion. Our study offers a theoretical framework to explain the emergence of local cell contraction pulses and their modulation by biochemical and mechanical signals.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Nocodazole, ≥99% (TLC), powder
Sigma-Aldrich
Biotin 3-sulfo-N-hydroxysuccinimide ester sodium salt, ≥90% (TLC), powder
Sigma-Aldrich
Collagen from calf skin, Bornstein and Traub Type I, (0.1% solution in 0.1 M acetic acid), aseptically processed, BioReagent, suitable for cell culture
Supelco
GC Stationary Phase, phase 1,2,3-tris(2-Cyanoethoxy)propane (TCEP), bottle of 50 g