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Subcellular optogenetic inhibition of G proteins generates signaling gradients and cell migration

Cells sense gradients of extracellular cues and generate polarized responses such as cell migration and neurite initiation. There is static information on the intracellular signaling molecules involved in these responses, but how they dynamically orchestrate polarized cell behaviors is not well unde...

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Detalles Bibliográficos
Autores principales: O'Neill, Patrick R., Gautam, N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4116304/
https://www.ncbi.nlm.nih.gov/pubmed/24920824
http://dx.doi.org/10.1091/mbc.E14-04-0870
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author O'Neill, Patrick R.
Gautam, N.
author_facet O'Neill, Patrick R.
Gautam, N.
author_sort O'Neill, Patrick R.
collection PubMed
description Cells sense gradients of extracellular cues and generate polarized responses such as cell migration and neurite initiation. There is static information on the intracellular signaling molecules involved in these responses, but how they dynamically orchestrate polarized cell behaviors is not well understood. A limitation has been the lack of methods to exert spatial and temporal control over specific signaling molecules inside a living cell. Here we introduce optogenetic tools that act downstream of native G protein–coupled receptor (GPCRs) and provide direct control over the activity of endogenous heterotrimeric G protein subunits. Light-triggered recruitment of a truncated regulator of G protein signaling (RGS) protein or a Gβγ-sequestering domain to a selected region on the plasma membrane results in localized inhibition of G protein signaling. In immune cells exposed to spatially uniform chemoattractants, these optogenetic tools allow us to create reversible gradients of signaling activity. Migratory responses generated by this approach show that a gradient of active G protein αi and βγ subunits is sufficient to generate directed cell migration. They also provide the most direct evidence so for a global inhibition pathway triggered by Gi signaling in directional sensing and adaptation. These optogenetic tools can be applied to interrogate the mechanistic basis of other GPCR-modulated cellular functions.
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spelling pubmed-41163042014-10-16 Subcellular optogenetic inhibition of G proteins generates signaling gradients and cell migration O'Neill, Patrick R. Gautam, N. Mol Biol Cell Articles Cells sense gradients of extracellular cues and generate polarized responses such as cell migration and neurite initiation. There is static information on the intracellular signaling molecules involved in these responses, but how they dynamically orchestrate polarized cell behaviors is not well understood. A limitation has been the lack of methods to exert spatial and temporal control over specific signaling molecules inside a living cell. Here we introduce optogenetic tools that act downstream of native G protein–coupled receptor (GPCRs) and provide direct control over the activity of endogenous heterotrimeric G protein subunits. Light-triggered recruitment of a truncated regulator of G protein signaling (RGS) protein or a Gβγ-sequestering domain to a selected region on the plasma membrane results in localized inhibition of G protein signaling. In immune cells exposed to spatially uniform chemoattractants, these optogenetic tools allow us to create reversible gradients of signaling activity. Migratory responses generated by this approach show that a gradient of active G protein αi and βγ subunits is sufficient to generate directed cell migration. They also provide the most direct evidence so for a global inhibition pathway triggered by Gi signaling in directional sensing and adaptation. These optogenetic tools can be applied to interrogate the mechanistic basis of other GPCR-modulated cellular functions. The American Society for Cell Biology 2014-08-01 /pmc/articles/PMC4116304/ /pubmed/24920824 http://dx.doi.org/10.1091/mbc.E14-04-0870 Text en © 2014 O'Neill and Gautam. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.
spellingShingle Articles
O'Neill, Patrick R.
Gautam, N.
Subcellular optogenetic inhibition of G proteins generates signaling gradients and cell migration
title Subcellular optogenetic inhibition of G proteins generates signaling gradients and cell migration
title_full Subcellular optogenetic inhibition of G proteins generates signaling gradients and cell migration
title_fullStr Subcellular optogenetic inhibition of G proteins generates signaling gradients and cell migration
title_full_unstemmed Subcellular optogenetic inhibition of G proteins generates signaling gradients and cell migration
title_short Subcellular optogenetic inhibition of G proteins generates signaling gradients and cell migration
title_sort subcellular optogenetic inhibition of g proteins generates signaling gradients and cell migration
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4116304/
https://www.ncbi.nlm.nih.gov/pubmed/24920824
http://dx.doi.org/10.1091/mbc.E14-04-0870
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