A genetic library screen for signaling proteins that interact with phosphorylated T cell costimulatory receptors

Xingxing Zang, P'ng Loke, Jayon Kim, Kathleen Wojnoonski, Leonard Kusdra, James P. Allison

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)841-845
Number of pages5
JournalGenomics
Volume88
Issue number6
DOIs
StatePublished - Dec 2006
Externally publishedYes

Fingerprint

Costimulatory and Inhibitory T-Cell Receptors
Inducible T-Cell Co-Stimulator Protein
Libraries
Phosphatidylinositol 3-Kinase
Allergy and Immunology
T-Lymphocytes
Proteins
Two-Hybrid System Techniques
Post Translational Protein Processing
Lymphocyte Activation
Gene Library
Autoimmune Diseases
Yeasts
Infection
Research
Neoplasms

Keywords

  • Costimulation
  • Functional genomics
  • Genetic screening
  • Inducible costimulator
  • Phosphatidylinositol 3-kinase
  • Phosphorylation
  • T lymphocytes

ASJC Scopus subject areas

  • Genetics

Cite this

A genetic library screen for signaling proteins that interact with phosphorylated T cell costimulatory receptors. / Zang, Xingxing; Loke, P'ng; Kim, Jayon; Wojnoonski, Kathleen; Kusdra, Leonard; Allison, James P.

In: Genomics, Vol. 88, No. 6, 12.2006, p. 841-845.

Research output: Contribution to journalArticle

Zang, Xingxing ; Loke, P'ng ; Kim, Jayon ; Wojnoonski, Kathleen ; Kusdra, Leonard ; Allison, James P. / A genetic library screen for signaling proteins that interact with phosphorylated T cell costimulatory receptors. In: Genomics. 2006 ; Vol. 88, No. 6. pp. 841-845.
@article{c88addd34be84ccf9a34ecd90a8bba57,
title = "A genetic library screen for signaling proteins that interact with phosphorylated T cell costimulatory receptors",
abstract = "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.",
keywords = "Costimulation, Functional genomics, Genetic screening, Inducible costimulator, Phosphatidylinositol 3-kinase, Phosphorylation, T lymphocytes",
author = "Xingxing Zang and P'ng Loke and Jayon Kim and Kathleen Wojnoonski and Leonard Kusdra and Allison, {James P.}",
year = "2006",
month = "12",
doi = "10.1016/j.ygeno.2006.08.012",
language = "English (US)",
volume = "88",
pages = "841--845",
journal = "Genomics",
issn = "0888-7543",
publisher = "Academic Press Inc.",
number = "6",

}

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.

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

UR - http://www.scopus.com/inward/record.url?scp=33750958715&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33750958715&partnerID=8YFLogxK

U2 - 10.1016/j.ygeno.2006.08.012

DO - 10.1016/j.ygeno.2006.08.012

M3 - Article

VL - 88

SP - 841

EP - 845

JO - Genomics

JF - Genomics

SN - 0888-7543

IS - 6

ER -