Abstract
The control of protein-protein interactions is a fundamental aspect of cell regulation. Here we describe a new approach to detect the interaction of two proteins in vivo. By this method, one binding partner is an integral membrane protein whereas the other is soluble but fused to a G-protein gamma-subunit. If the binding partners interact, G-protein signaling is disrupted. We demonstrate interaction between known binding partners, syntaxin 1a with neuronal Sec1 (nSec1), and the fibroblast-derived growth factor receptor 3 (FGFR3) with SNT-1. In addition, we describe a genetic screen to identify nSec1 mutants that are expressed normally, but are no longer able to bind to syntaxin 1a. This provides a convenient method to study interactions of integral membrane proteins, a class of molecules that has been difficult to study by existing biochemical or genetic methods.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adaptor Proteins, Signal Transducing
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Antigens, Surface / genetics
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Antigens, Surface / metabolism
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Cell Membrane / chemistry
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Cell Membrane / metabolism*
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Cytoplasm / chemistry
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Cytoplasm / metabolism
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GTP-Binding Protein gamma Subunits*
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Genes, Reporter / genetics
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Heterotrimeric GTP-Binding Proteins / genetics*
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Heterotrimeric GTP-Binding Proteins / metabolism*
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Munc18 Proteins
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Mutation / genetics
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Nerve Tissue Proteins / genetics
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Nerve Tissue Proteins / metabolism
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Pheromones / pharmacology
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Phosphoproteins / genetics
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Phosphoproteins / metabolism
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Protein Binding / drug effects
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Protein Transport
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Protein-Tyrosine Kinases*
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Receptor, Fibroblast Growth Factor, Type 3
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Receptors, Fibroblast Growth Factor / genetics
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Receptors, Fibroblast Growth Factor / metabolism
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Saccharomyces cerevisiae / cytology
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Saccharomyces cerevisiae / drug effects
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae Proteins*
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Signal Transduction / drug effects
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Solubility
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Syntaxin 1
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Two-Hybrid System Techniques*
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Vesicular Transport Proteins*
Substances
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Adaptor Proteins, Signal Transducing
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Antigens, Surface
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FRS2 protein, human
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GTP-Binding Protein gamma Subunits
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Membrane Proteins
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Munc18 Proteins
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Nerve Tissue Proteins
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Pheromones
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Phosphoproteins
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Receptors, Fibroblast Growth Factor
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Recombinant Fusion Proteins
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STE18 protein, S cerevisiae
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STX1A protein, human
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Saccharomyces cerevisiae Proteins
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Syntaxin 1
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Vesicular Transport Proteins
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Protein-Tyrosine Kinases
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Receptor, Fibroblast Growth Factor, Type 3
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Heterotrimeric GTP-Binding Proteins