A combined binary interaction and phenotypic map of C. elegans cell polarity proteins

Nature Cell Biology
Thijs KoormanMike Boxem

Abstract

The establishment of cell polarity is an essential process for the development of multicellular organisms and the functioning of cells and tissues. Here, we combine large-scale protein interaction mapping with systematic phenotypic profiling to study the network of physical interactions that underlies polarity establishment and maintenance in the nematode Caenorhabditis elegans. Using a fragment-based yeast two-hybrid strategy, we identified 439 interactions between 296 proteins, as well as the protein regions that mediate these interactions. Phenotypic profiling of the network resulted in the identification of 100 physically interacting protein pairs for which RNAi-mediated depletion caused a defect in the same polarity-related process. We demonstrate the predictive capabilities of the network by showing that the physical interaction between the RhoGAP PAC-1 and PAR-6 is required for radial polarization of the C. elegans embryo. Our network represents a valuable resource of candidate interactions that can be used to further our insight into cell polarization.

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Citations

Aug 9, 2016·Annual Review of Cell and Developmental Biology·Yelena Bernadskaya, Lionel Christiaen
Nov 3, 2016·Molecular & Cellular Proteomics : MCP·Sam LievensJan Tavernier
Jan 26, 2018·Wiley Interdisciplinary Reviews. Developmental Biology·John Isaac Murray

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Methods Mentioned

BETA
two-hybrid
co-affinity purification
co-affinity purifications
Y2H
PCR
two hybrid
transgenic
co-affinity
FCS

Software Mentioned

Gene Ontology ( HRSS )
AxioVision
phred
MetaMorph
WormNet
Cluster
MAPPIT
Adobe Photoshop
Zeiss Axiovision
BLAST

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