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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

Supelco
GC Stationary Phase, phase 1,2,3-tris(2-Cyanoethoxy)propane (TCEP), bottle of 50 g
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