Synthetic mechanobiology: engineering cellular force generation and signaling

Hughes, Jasmine H.; Kumar, Sanjay

doi:10.1016/j.copbio.2016.03.004

Cited by 14 publications

(13 citation statements)

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“…, 1997 ). Importantly, doxycycline induction allows titration of gene expression over a continuous range, which in turn enables elucidation of quantitative relationships between expression and mechanobiological phenotype in a manner not possible with pharmacological inhibition or transient plasmid overexpression ( MacKay et al., 2012 , 2014 ; MacKay and Kumar, 2014 ; Hughes and Kumar, 2016 ). Understanding this dose-response relationship is an important experimental design consideration given that the relationship between myosin activation and mechanobiological phenotype is often highly nonlinear ( MacKay and Kumar, 2014 ; Rape et al.…”

Section: Resultsmentioning

confidence: 99%

Activation of ROCK and MLCK tunes regional stress fiber formation and mechanics via preferential myosin light chain phosphorylation

Kassianidou

Hughes

Kumar

2017

MBoC

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MBoC | ARTICLE Activation of ROCK and MLCK tunes regional stress fiber formation and mechanics via preferential myosin light chain phosphorylation ABSTRACT The assembly and mechanics of actomyosin stress fibers (SFs) depend on myosin regulatory light chain (RLC) phosphorylation, which is driven by myosin light chain kinase (MLCK) and Rho-associated kinase (ROCK). Although previous work suggests that MLCK and ROCK control distinct pools of cellular SFs, it remains unclear how these kinases differ in their regulation of RLC phosphorylation or how phosphorylation influences individual SF mechanics. Here, we combine genetic approaches with biophysical tools to explore relationships between kinase activity, RLC phosphorylation, SF localization, and SF mechanics. We show that graded MLCK overexpression increases RLC monophosphorylation (p-RLC) in a graded manner and that this p-RLC localizes to peripheral SFs. Conversely, graded ROCK overexpression preferentially increases RLC diphosphorylation (pp-RLC), with pp-RLC localizing to central SFs. Interrogation of single SFs with subcellular laser ablation reveals that MLCK and ROCK quantitatively regulate the viscoelastic properties of peripheral and central SFs, respectively. The effects of MLCK and ROCK on single-SF mechanics may be correspondingly phenocopied by overexpression of mono-and diphosphomimetic RLC mutants. Our results point to a model in which MLCK and ROCK regulate peripheral and central SF viscoelastic properties through mono-and diphosphorylation of RLC, offering new quantitative connections between kinase activity, RLC phosphorylation, and SF viscoelasticity.

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Section: Resultsmentioning

confidence: 99%

Activation of ROCK and MLCK tunes regional stress fiber formation and mechanics via preferential myosin light chain phosphorylation

Kassianidou

Hughes

Kumar

2017

MBoC

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“…Compartmentalizing the location of proteins in the cytosol can be effectively altered though the application of subcellular forces. Mechanically manipulating the position of proteins can be controlled through endocytosed magnetic nanoparticles within magnetic field gradients (Pan et al, 2012 ; Bonnemay et al, 2013 ; Etoc et al, 2013 , 2015 ; Kunze et al, 2015 ; Hughes and Kumar, 2016 ; Ducasse et al, 2017 ; Liße et al, 2017 ; Monzel et al, 2017 ). The force range to establish a specific protein gradient, however, should leave the tension at the cell membrane at a homeostatic level.…”

Section: Compartmentalizing Intracellular Proteinsmentioning

confidence: 99%

Force-Mediating Magnetic Nanoparticles to Engineer Neuronal Cell Function

Gahl

Kunze

2018

Front. Neurosci.

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Cellular processes like membrane deformation, cell migration, and transport of organelles are sensitive to mechanical forces. Technically, these cellular processes can be manipulated through operating forces at a spatial precision in the range of nanometers up to a few micrometers through chaperoning force-mediating nanoparticles in electrical, magnetic, or optical field gradients. But which force-mediating tool is more suitable to manipulate cell migration, and which, to manipulate cell signaling? We review here the differences in forces sensation to control and engineer cellular processes inside and outside the cell, with a special focus on neuronal cells. In addition, we discuss technical details and limitations of different force-mediating approaches and highlight recent advancements of nanomagnetics in cell organization, communication, signaling, and intracellular trafficking. Finally, we give suggestions about how force-mediating nanoparticles can be used to our advantage in next-generation neurotherapeutic devices.

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“…Finally, the transposition of our strategy into living cells will be powerful to target and manipulate specific proteins and cellular organelles by combining chemically induced dimerization and magnetic manipulation 23 , 63 , 65 . A fully-genetically encoded strategy, avoiding in vitro biomineralisation and injection into cells, will require novel approaches to catalyse enhanced magnetic phases within cells.…”

Section: Discussionmentioning

confidence: 99%

Programmed Self-Assembly of a Biochemical and Magnetic Scaffold to Trigger and Manipulate Microtubule Structures

Ducasse

Wang

Navarro

et al. 2017

Sci Rep

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Artificial bio-based scaffolds offer broad applications in bioinspired chemistry, nanomedicine, and material science. One current challenge is to understand how the programmed self-assembly of biomolecules at the nanometre level can dictate the emergence of new functional properties at the mesoscopic scale. Here we report a general approach to design genetically encoded protein-based scaffolds with modular biochemical and magnetic functions. By combining chemically induced dimerization strategies and biomineralisation, we engineered ferritin nanocages to nucleate and manipulate microtubule structures upon magnetic actuation. Triggering the self-assembly of engineered ferritins into micrometric scaffolds mimics the function of centrosomes, the microtubule organizing centres of cells, and provides unique magnetic and self-organizing properties. We anticipate that our approach could be transposed to control various biological processes and extend to broader applications in biotechnology or material chemistry.

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Synthetic mechanobiology: engineering cellular force generation and signaling

Cited by 14 publications

References 57 publications

Activation of ROCK and MLCK tunes regional stress fiber formation and mechanics via preferential myosin light chain phosphorylation

Activation of ROCK and MLCK tunes regional stress fiber formation and mechanics via preferential myosin light chain phosphorylation

Force-Mediating Magnetic Nanoparticles to Engineer Neuronal Cell Function

Programmed Self-Assembly of a Biochemical and Magnetic Scaffold to Trigger and Manipulate Microtubule Structures

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