Differential Contributions of Nonmuscle Myosin II Isoforms and Functional Domains to Stress Fiber Mechanics

Scientific Reports
Ching-Wei Chang, Sanjay Kumar

Abstract

While is widely acknowledged that nonmuscle myosin II (NMMII) enables stress fibers (SFs) to generate traction forces against the extracellular matrix, little is known about how specific NMMII isoforms and functional domains contribute to SF mechanics. Here we combine biophotonic and genetic approaches to address these open questions. First, we suppress the NMMII isoforms MIIA and MIIB and apply femtosecond laser nanosurgery to ablate and investigate the viscoelastic retraction of individual SFs. SF retraction dynamics associated with MIIA and MIIB suppression qualitatively phenocopy our earlier measurements in the setting of Rho kinase (ROCK) and myosin light chain kinase (MLCK) inhibition, respectively. Furthermore, fluorescence imaging and photobleaching recovery reveal that MIIA and MIIB are enriched in and more stably localize to ROCK- and MLCK-controlled central and peripheral SFs, respectively. Additional domain-mapping studies surprisingly reveal that deletion of the head domain speeds SF retraction, which we ascribe to reduced drag from actomyosin crosslinking and frictional losses. We propose a model in which ROCK/MIIA and MLCK/MIIB functionally regulate common pools of SFs, with MIIA crosslinking and motor functions ...Continue Reading

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Citations

Feb 23, 2018·Molecular Biology of the Cell·Masahiro KuraganoMasayuki Takahashi
Jun 22, 2018·Molecular Biology of the Cell·Stacey LeeSanjay Kumar
Nov 24, 2018·Biomolecules·Beth S Lee
Sep 17, 2016·F1000Research·Stacey Lee, Sanjay Kumar
Sep 6, 2019·ELife·Mélina L HeuzéBenoit Ladoux
Feb 19, 2017·Proceedings of the National Academy of Sciences of the United States of America·Elena KassianidouSanjay Kumar
Mar 20, 2018·Biochemical and Biophysical Research Communications·Maria S Shutova, Tatyana M Svitkina

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Methods Mentioned

BETA
fluorescence recovery after photobleaching
immunoprecipitation
targeted mutations
transfection

Software Mentioned

MathWorks
MATLAB

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