TY - JOUR
T1 - A genetic library screen for signaling proteins that interact with phosphorylated T cell costimulatory receptors
AU - Zang, Xingxing
AU - Loke, P'ng
AU - Kim, Jayon
AU - Wojnoonski, Kathleen
AU - Kusdra, Leonard
AU - Allison, James P.
N1 - Funding Information:
We thank Tsvetelina Hoang, Pete Savage, and Alejandro Sepulveda for critically reading the manuscript. This work was supported by the National Institutes of Health (2R01CA040041-19) and the Howard Hughes Medical Institute. X.Z. is a Fellow of the Cancer Research Institute, P.L. is the recipient of a Wellcome International Research Fellowship, and J.P.A. is an Investigator of the Howard Hughes Medical Institute.
PY - 2006/12
Y1 - 2006/12
N2 - In the past decade, the fundamental importance and therapeutic potential of costimulatory signals for lymphocyte activation have spurred a large amount of work in immunology, infection, cancer, autoimmune diseases, etc. However, the mechanisms behind T cell costimulation remain unclear, partly due to the lack of suitable techniques. There is an urgent need for functional genomic research to develop comprehensive approaches to direct identification of protein-protein interactions that are dependent on the posttranslational modification of one component of the complex, particularly in the field of T cell immunology. Using inducible costimulator (ICOS) as a model, we failed to find any proteins that associated with the cytoplasmic tail of ICOS by the yeast two-hybrid approach. Therefore, we have developed a new yeast three-hybrid system that facilitates the rapid screening of cDNA libraries to find signaling molecules that interact with phosphorylated T cell costimulatory receptors. We demonstrate the utility of this technique to detect the interaction between ICOS and the p85 regulatory subunit of phosphatidylinositol 3-kinase (PI3K). The p85 unit of PI3K is the only signaling molecule identified so far that interacts with ICOS. This system may be of great help in dissecting the mechanisms of T cell costimulation and could be applied to other receptors.
AB - In the past decade, the fundamental importance and therapeutic potential of costimulatory signals for lymphocyte activation have spurred a large amount of work in immunology, infection, cancer, autoimmune diseases, etc. However, the mechanisms behind T cell costimulation remain unclear, partly due to the lack of suitable techniques. There is an urgent need for functional genomic research to develop comprehensive approaches to direct identification of protein-protein interactions that are dependent on the posttranslational modification of one component of the complex, particularly in the field of T cell immunology. Using inducible costimulator (ICOS) as a model, we failed to find any proteins that associated with the cytoplasmic tail of ICOS by the yeast two-hybrid approach. Therefore, we have developed a new yeast three-hybrid system that facilitates the rapid screening of cDNA libraries to find signaling molecules that interact with phosphorylated T cell costimulatory receptors. We demonstrate the utility of this technique to detect the interaction between ICOS and the p85 regulatory subunit of phosphatidylinositol 3-kinase (PI3K). The p85 unit of PI3K is the only signaling molecule identified so far that interacts with ICOS. This system may be of great help in dissecting the mechanisms of T cell costimulation and could be applied to other receptors.
KW - Costimulation
KW - Functional genomics
KW - Genetic screening
KW - Inducible costimulator
KW - Phosphatidylinositol 3-kinase
KW - Phosphorylation
KW - T lymphocytes
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U2 - 10.1016/j.ygeno.2006.08.012
DO - 10.1016/j.ygeno.2006.08.012
M3 - Article
C2 - 17014982
AN - SCOPUS:33750958715
SN - 0888-7543
VL - 88
SP - 841
EP - 845
JO - Genomics
JF - Genomics
IS - 6
ER -