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Dev Cell. 2017 Aug 21;42(4):416-434.e11. doi: 10.1016/j.devcel.2017.07.024.

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

Author information

1
Department of Biology and Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. Electronic address: daniel.dickinson@austin.utexas.edu.
2
Department of Cell Physiology and Metabolism, University of Geneva Medical Faculty, 1, rue Michel Servet, 1211 Geneva, Switzerland.
3
Institut Jacques Monod, Cell Cycle and Development Team, Centre National de la Recherche Scientifique and University of Paris Diderot and Sorbonne Paris Cité UMR7592, Paris 75013, France.
4
Department of Biology and Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

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:

C. elegans; PAR proteins; PAR-3; PLK-1; SiMPull; cell polarity; cortical flow; sc-SiMPull; single-cell biochemistry

PMID:
28829947
PMCID:
PMC5575849
DOI:
10.1016/j.devcel.2017.07.024
[Indexed for MEDLINE]
Free PMC Article

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