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The genetic design of signaling cascades to record receptor activation

Author:
Barnea, Gilad, Strapps, Walter, Herrada, Gilles, Berman, Yemiliya, Ong, Jane, Kloss, Brian, Axel, Richard, Lee, Kevin J.
Source:
Proceedings of the National Academy of Sciences of the United States of America 2008 v.105 no.1 pp. 64-69
ISSN:
0027-8424
Subject:
proteinases, receptor protein-tyrosine kinase, G-protein coupled receptors, inflammation, signal transduction, steroid hormone receptors, reporter genes
Abstract:
We have developed an experimental strategy to monitor protein interactions in a cell with a high degree of selectivity and sensitivity. A transcription factor is tethered to a membrane-bound receptor with a linker that contains a cleavage site for a specific protease. Activation of the receptor recruits a signaling protein fused to the protease that then cleaves and releases the transcription factor to activate reporter genes in the nucleus. This strategy converts a transient interaction into a stable and amplifiable reporter gene signal to record the activation of a receptor without interference from endogenous signaling pathways. We have developed this assay for three classes of receptors: G protein-coupled receptors, receptor tyrosine kinases, and steroid hormone receptors. Finally, we use the assay to identify a ligand for the orphan receptor GPR1, suggesting a role for this receptor in the regulation of inflammation.
Agid:
2360190