Journal article
Multipoint Binding of the SLP-76 SH2 Domain to ADAP Is Critical for Oligomerization of SLP-76 Signaling Complexes in Stimulated T Cells
Molecular and cellular biology, Vol.33(21), pp.4140-4151
11/2013
DOI: 10.1128/MCB.00410-13
PMCID: PMC3811887
PMID: 23979596
Abstract
The adapter molecules SLP-76 and LAT play central roles in T cell activation by recruiting enzymes and other adapters into multiprotein complexes that coordinate highly regulated signal transduction pathways. While many of the associated proteins have been characterized, less is known concerning the mechanisms of assembly for these dynamic and potentially heterogeneous signaling complexes. Following T cell receptor (TCR) stimulation, SLP-76 is found in structures called microclusters, which contain many signaling complexes. Previous studies showed that a mutation to the SLP-76 C-terminal SH2 domain nearly abolished SLP-76 microclusters, suggesting that the SH2 domain facilitates incorporation of signaling complexes into microclusters. S. C. Bunnell, A. L. Singer, D. I. Hong, B. H. Jacque, M. S. Jordan, M. C. Seminario, V. A. Barr, G. A. Koretzky, and L. E. Samelson, Mol. Cell. Biol., 26:7155–7166, 2006). Using biophysical methods, we demonstrate that the adapter, ADAP, contains three binding sites for SLP-76, and that multipoint binding to ADAP fragments oligomerizes the SLP-76 SH2 domain
in vitro
. These results were complemented with confocal imaging and functional studies of cells expressing ADAP with various mutations. Our results demonstrate that all three binding sites are critical for SLP-76 microcluster assembly, but any combination of two sites will partially induce microclusters. These data support a model whereby multipoint binding of SLP-76 to ADAP facilitates the assembly of SLP-76 microclusters. This model has implications for the regulation of SLP-76 and LAT microclusters and, as a result, T cell signaling.
Details
- Title: Subtitle
- Multipoint Binding of the SLP-76 SH2 Domain to ADAP Is Critical for Oligomerization of SLP-76 Signaling Complexes in Stimulated T Cells
- Creators
- Nathan P Coussens - Laboratory of Cellular and Molecular BiologyRyo Hayashi - Laboratory of Cell BiologyPatrick H Brown - Biomedical Engineering and Physical Sciences Shared ResourceLakshmi Balagopalan - Laboratory of Cellular and Molecular BiologyAndrea Balbo - Biomedical Engineering and Physical Sciences Shared ResourceItoro Akpan - Laboratory of Cellular and Molecular BiologyJon C. D Houtman - Laboratory of Cellular and Molecular BiologyValarie A Barr - Laboratory of Cellular and Molecular BiologyPeter Schuck - Dynamics of Macromolecular Assembly SectionEttore Appella - Laboratory of Cell BiologyLawrence E Samelson - Laboratory of Cellular and Molecular Biology
- Resource Type
- Journal article
- Publication Details
- Molecular and cellular biology, Vol.33(21), pp.4140-4151
- DOI
- 10.1128/MCB.00410-13
- PMID
- 23979596
- PMCID
- PMC3811887
- NLM abbreviation
- Mol Cell Biol
- ISSN
- 0270-7306
- eISSN
- 1098-5549
- Publisher
- American Society for Microbiology
- Language
- English
- Date published
- 11/2013
- Academic Unit
- Microbiology and Immunology; Internal Medicine
- Record Identifier
- 9984094330402771
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