The Roles of Signaling in Cytoskeletal Changes, Random Movement, Direction-Sensing and Polarization of Eukaryotic Cells

Cheng, Yuanna; Félix, Bryan; Othmer, Hans G.

doi:10.3390/cells9061437

Cited by 21 publications

(20 citation statements)

References 190 publications

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“…Alterations of the levels of second messengers such as cAMP or Ca 2+ often precede any detectable changes in cytoskeletal dynamics, a key determinant of the cell shape ( 92 , 93 , 94 , 95 , 96 , 97 ). Activation of GPCRs from all coupling classes gives shape to cells and enables distinctly different changes of cellular morphology in a G protein subfamily–specific manner ( 98 , 99 ).…”

Section: Resultsmentioning

confidence: 99%

An experimental strategy to probe Gq contribution to signal transduction in living cells

Patt

Alenfelder

Pfeil

et al. 2021

Journal of Biological Chemistry

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Heterotrimeric G protein subunits Gαq and Gα11 are inhibited by two cyclic depsipeptides, FR900359 (FR) and YM-254890 (YM), both of which are being used widely to implicate Gq/11 proteins in the regulation of diverse biological processes. An emerging major research question therefore is whether the cellular effects of both inhibitors are on-target, that is, mediated via specific inhibition of Gq/11 proteins, or off-target, that is, the result of nonspecific interactions with other proteins. Here we introduce a versatile experimental strategy to discriminate between these possibilities. We developed a Gαq variant with preserved catalytic activity, but refractory to FR/YM inhibition. A minimum of two amino acid changes were required and sufficient to achieve complete inhibitor resistance. We characterized the novel mutant in HEK293 cells depleted by CRISPR–Cas9 of endogenous Gαq and Gα11 to ensure precise control over the Gα-dependent cellular signaling route. Using a battery of cellular outcomes with known and concealed Gq contribution, we found that FR/YM specifically inhibited cellular signals after Gαq introduction via transient transfection. Conversely, both inhibitors were inert across all assays in cells expressing the drug-resistant variant. These findings eliminate the possibility that inhibition of non-Gq proteins contributes to the cellular effects of the two depsipeptides. We conclude that combined application of FR or YM along with the drug-resistant Gαq variant is a powerful in vitro strategy to discern on-target Gq against off-target non-Gq action. Consequently, it should be of high value for uncovering Gq input to complex biological processes with high accuracy and the requisite specificity.

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Section: Resultsmentioning

confidence: 99%

An experimental strategy to probe Gq contribution to signal transduction in living cells

Patt

Alenfelder

Pfeil

et al. 2021

Journal of Biological Chemistry

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“…Conversely, the Ras GTPase activating proteins RasGAP2 and RasGAP3, as well as phosphatidylinositol bisphosphate (PIP2) show higher concentrations at the trailing edge of the cell and render Ras in an inactive state (Ras-GDP), reducing actin polymerization (Li et al 2018). The activation of Ras at the leading edge of the cell thus causes directed migration of amoeboid cells (Cai et al 2010;Cheng, Felix, and Othmer 2020;Rappel and Edelstein-Keshet 2017).…”

Section: Introductionmentioning

confidence: 99%

A chemorepellent inhibits local Ras activation to inhibit pseudopod formation to bias cell movement away from the chemorepellent

Kirolos

Gomer

2022

MBoC

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The ability of cells to sense chemical gradients is essential during development, morphogenesis, and immune responses. Although much is known about chemoattraction, chemorepulsion remains poorly understood. Proliferating Dictyostelium cells secrete a chemorepellent protein called AprA. AprA prevents pseudopod formation at the region of the cell closest to the source of AprA, causing the random movement of cells to be biased away from the AprA. Activation of Ras proteins in a localized sector of a cell cortex helps to induce pseudopod formation, and Ras proteins are needed for AprA chemorepulsion. Here we show that AprA locally inhibits Ras cortical activation through the G protein-coupled receptor GrlH, the G protein subunits Gβ and Gα8, Ras protein RasG, protein kinase B, the p-21 activated kinase PakD, and the extracellular signal-regulated kinase Erk1. Diffusion calculations and experiments indicate that in a colony of cells, high extracellular concentrations of AprA in the center can globally inhibit Ras activation, while a gradient of AprA that naturally forms at the edge of the colony allow cells to activate Ras at sectors of the cell other than the sector of the cell closest to the center of the colony, effectively inducing both repulsion from the colony and cell differentiation. Together, these results suggest that a pathway that inhibits local Ras activation can mediate chemorepulsion. [Media: see text] [Media: see text] [Media: see text] [Media: see text] [Media: see text] [Media: see text]

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“…Moreover, the regulation of cytoskeleton and metabolic signaling pathways were involved in tissue regeneration for the cell movement, growth, and proliferation [ 33 , 34 ]. Studies showed that the modification of lipid metabolism is needed in liver and axon regeneration [ 35 , 36 ].…”

Section: Resultsmentioning

confidence: 99%

A Brief Analysis of Proteomic Profile Changes during Zebrafish Regeneration

Ulhaq

Tse

2021

Biomolecules

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Unlike mammals, zebrafish are capable to regenerate many of their organs, however, the response of tissue damage varies across tissues. Understanding the molecular mechanism behind the robust regenerative capacity in a model organism may help to identify and develop novel treatment strategies for mammals (including humans). Hence, we systematically analyzed the current literature on the proteome profile collected from different regenerated zebrafish tissues. Our analyses underlining that several proteins and protein families responsible as a component of cytoskeleton and structure, protein synthesis and degradation, cell cycle control, and energy metabolism were frequently identified. Moreover, target proteins responsible for the initiation of the regeneration process, such as inflammation and immune response were less frequently detected. This highlights the limitation of previous proteomic analysis and suggested a more sensitive modern proteomics analysis is needed to unfold the mechanism. This brief report provides a list of target proteins with predicted functions that could be useful for further biological studies.

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The Roles of Signaling in Cytoskeletal Changes, Random Movement, Direction-Sensing and Polarization of Eukaryotic Cells

Cited by 21 publications

References 190 publications

An experimental strategy to probe Gq contribution to signal transduction in living cells

An experimental strategy to probe Gq contribution to signal transduction in living cells

A chemorepellent inhibits local Ras activation to inhibit pseudopod formation to bias cell movement away from the chemorepellent

A Brief Analysis of Proteomic Profile Changes during Zebrafish Regeneration

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