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A Single-Cell Biochemistry Approach Reveals PAR Complex Dynamics during Cell Polarization

Dickinson, Daniel J
Pintard, Lionel
Goldstein, Bob
Published in Developmental cell. 2017, vol. 42, no. 4, p. 416-434.e11
Abstract Regulated protein-protein interactions are critical for cell signaling, differentiation, and development. For the study of dynamic regulation of protein interactions in vivo, there is a need for techniques that can yield time-resolved information and probe multiple protein binding partners simultaneously, using small amounts of starting material. Here we describe a single-cell protein interaction assay. Single-cell lysates are generated at defined time points and analyzed using single-molecule pull-down, yielding information about dynamic protein complex regulation in vivo. We established the utility of this approach by studying PAR polarity proteins, which mediate polarization of many animal cell types. We uncovered striking regulation of PAR complex composition and stoichiometry during Caenorhabditis elegans zygote polarization, which takes place in less than 20 min. PAR complex dynamics are linked to the cell cycle by Polo-like kinase 1 and govern the movement of PAR proteins to establish polarity. Our results demonstrate an approach to study dynamic biochemical events in vivo.
Keywords AnimalsCaenorhabditis elegans/cytology/metabolismCaenorhabditis elegans Proteins/metabolismCell PolarityImmunoprecipitation/methodsProtein BindingProtein MultimerizationProtein-Serine-Threonine Kinases/metabolismSingle-Cell Analysis/methodsZygote/cytology/metabolism
PMID: 28829947
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Research group La division asymétrique (847)
Swiss National Science Foundation: 31003A_138501
Swiss National Science Foundation: 31003A_156013
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DICKINSON, Daniel J et al. A Single-Cell Biochemistry Approach Reveals PAR Complex Dynamics during Cell Polarization. In: Developmental Cell, 2017, vol. 42, n° 4, p. 416-434.e11. doi: 10.1016/j.devcel.2017.07.024 https://archive-ouverte.unige.ch/unige:112242

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Deposited on : 2018-12-12

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