Functional Proteomics Mapping of a Human Signaling Pathway
Abstract
Access to the human genome facilitates extensive functional proteomics studies. Here, we present an integrated approach combining large-scale protein interaction mapping, exploration of the interaction network, and cellular functional assays performed on newly identified proteins involved in a human signaling pathway. As a proof of principle, we studied the Smad signaling system, which is regulated by members of the transforming growth factor β (TGFβ) superfamily. We used two-hybrid screening to map Smad signaling protein–protein interactions and to establish a network of 755 interactions, involving 591 proteins, 179 of which were poorly or not annotated. The exploration of such complex interaction databases is improved by the use of PIMRider, a dedicated navigation tool accessible through the Web. The biological meaning of this network is illustrated by the presence of 18 known Smad-associated proteins. Functional assays performed in mammalian cells including siRNA knock-down experiments identified eight novel proteins involved in Smad signaling, thus validating this integrated functional proteomics approach.
Footnotes
[Supplemental material is available online at www.genome.org.]
Article and publication are at http://www.genome.org/cgi/doi/10.1101/gr.2334104.
↵1Corresponding author.E-MAILfcolland{at}hybrigenics.fr; FAX 33 1 5810 3846.
- Accepted April 29, 2004.
- Received January 5, 2004.
- Cold Spring Harbor Laboratory Press
