Differential association of CD45 isoforms with CD4 and CD8 regulates the actions of specific pools of p56lck tyrosine kinase in T cell antigen receptor signal transduction

Saffron Dornan, Zsolt Sebestyen, John Gamble, Peter Nagy, Andrea Bodnar, Lou Alldridge, Senam Doe, Nick Holmes, Lindsey K. Goff, Peter Beverley, J. Szöllősi, Denis R. Alexander

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

An investigation into the role of CD45 isoforms in T cell antigen receptor signal transduction was carried out by transfecting CD45-negative CD4+CD8+ HPB-ALL T cells with the CD45R0, CD45RBC, and CD45RABC isoforms. Fluorescence resonance energy transfer analysis showed that the CD45R0 isoform, but not the CD45RBC or CD45RABC isoforms, was found as homodimers and also preferentially associated with CD4 and CD8 at the cell-surface. A comparison was therefore made of T cell antigen receptor signaling between sub-clones expressing either CD45R0 or CD45RBC. Under basal conditions CD4-associated p56lck tyrosine kinase activity and cellular protein tyrosine phosphorylation levels were higher in the CD45R0+ than in the CD45RBC+ sub-clones. Upon CD3-CD4 ligation, TCR-ζ phosphorylation, ZAP-70 recruitment to the p21/p23 TCR-ζ phosphoisomers, ZAP-70 phosphorylation, as well as p56lck, c-Cbl and Slp-76 phosphorylation, were all markedly increased in CD45R0+ compared with CD45RBC+ cells. T cell antigen receptor (TCR) stimulation alone also promoted c-Cbl phosphorylation in CD45R0+ but not in CD45RBC+ cells. Our results are consistent with a model in which association of CD45R0 with CD4 generates a more active pool of CD4-associated p56lck kinase molecules. Upon CD3-CD4 co-ligation, the active p56lck increases the intensity of T cell antigen receptor signal transduction coupling by promoting TCR-ζ chain phosphorylation and ZAP-70 recruitment.

Original languageEnglish
Pages (from-to)1912-1918
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number3
DOIs
Publication statusPublished - Jan 18 2002

Fingerprint

Signal transduction
Phosphorylation
T-Cell Antigen Receptor
Protein-Tyrosine Kinases
Signal Transduction
Protein Isoforms
Association reactions
Ligation
Clone Cells
Fluorescence Resonance Energy Transfer
T-cells
Tyrosine
Phosphotransferases
T-Lymphocytes
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Differential association of CD45 isoforms with CD4 and CD8 regulates the actions of specific pools of p56lck tyrosine kinase in T cell antigen receptor signal transduction. / Dornan, Saffron; Sebestyen, Zsolt; Gamble, John; Nagy, Peter; Bodnar, Andrea; Alldridge, Lou; Doe, Senam; Holmes, Nick; Goff, Lindsey K.; Beverley, Peter; Szöllősi, J.; Alexander, Denis R.

In: Journal of Biological Chemistry, Vol. 277, No. 3, 18.01.2002, p. 1912-1918.

Research output: Contribution to journalArticle

Dornan, S, Sebestyen, Z, Gamble, J, Nagy, P, Bodnar, A, Alldridge, L, Doe, S, Holmes, N, Goff, LK, Beverley, P, Szöllősi, J & Alexander, DR 2002, 'Differential association of CD45 isoforms with CD4 and CD8 regulates the actions of specific pools of p56lck tyrosine kinase in T cell antigen receptor signal transduction', Journal of Biological Chemistry, vol. 277, no. 3, pp. 1912-1918. https://doi.org/10.1074/jbc.M108386200
Dornan, Saffron ; Sebestyen, Zsolt ; Gamble, John ; Nagy, Peter ; Bodnar, Andrea ; Alldridge, Lou ; Doe, Senam ; Holmes, Nick ; Goff, Lindsey K. ; Beverley, Peter ; Szöllősi, J. ; Alexander, Denis R. / Differential association of CD45 isoforms with CD4 and CD8 regulates the actions of specific pools of p56lck tyrosine kinase in T cell antigen receptor signal transduction. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 3. pp. 1912-1918.
@article{54709fcac9904ee2bb0a30664d992635,
title = "Differential association of CD45 isoforms with CD4 and CD8 regulates the actions of specific pools of p56lck tyrosine kinase in T cell antigen receptor signal transduction",
abstract = "An investigation into the role of CD45 isoforms in T cell antigen receptor signal transduction was carried out by transfecting CD45-negative CD4+CD8+ HPB-ALL T cells with the CD45R0, CD45RBC, and CD45RABC isoforms. Fluorescence resonance energy transfer analysis showed that the CD45R0 isoform, but not the CD45RBC or CD45RABC isoforms, was found as homodimers and also preferentially associated with CD4 and CD8 at the cell-surface. A comparison was therefore made of T cell antigen receptor signaling between sub-clones expressing either CD45R0 or CD45RBC. Under basal conditions CD4-associated p56lck tyrosine kinase activity and cellular protein tyrosine phosphorylation levels were higher in the CD45R0+ than in the CD45RBC+ sub-clones. Upon CD3-CD4 ligation, TCR-ζ phosphorylation, ZAP-70 recruitment to the p21/p23 TCR-ζ phosphoisomers, ZAP-70 phosphorylation, as well as p56lck, c-Cbl and Slp-76 phosphorylation, were all markedly increased in CD45R0+ compared with CD45RBC+ cells. T cell antigen receptor (TCR) stimulation alone also promoted c-Cbl phosphorylation in CD45R0+ but not in CD45RBC+ cells. Our results are consistent with a model in which association of CD45R0 with CD4 generates a more active pool of CD4-associated p56lck kinase molecules. Upon CD3-CD4 co-ligation, the active p56lck increases the intensity of T cell antigen receptor signal transduction coupling by promoting TCR-ζ chain phosphorylation and ZAP-70 recruitment.",
author = "Saffron Dornan and Zsolt Sebestyen and John Gamble and Peter Nagy and Andrea Bodnar and Lou Alldridge and Senam Doe and Nick Holmes and Goff, {Lindsey K.} and Peter Beverley and J. Sz{\"o}llősi and Alexander, {Denis R.}",
year = "2002",
month = "1",
day = "18",
doi = "10.1074/jbc.M108386200",
language = "English",
volume = "277",
pages = "1912--1918",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "3",

}

TY - JOUR

T1 - Differential association of CD45 isoforms with CD4 and CD8 regulates the actions of specific pools of p56lck tyrosine kinase in T cell antigen receptor signal transduction

AU - Dornan, Saffron

AU - Sebestyen, Zsolt

AU - Gamble, John

AU - Nagy, Peter

AU - Bodnar, Andrea

AU - Alldridge, Lou

AU - Doe, Senam

AU - Holmes, Nick

AU - Goff, Lindsey K.

AU - Beverley, Peter

AU - Szöllősi, J.

AU - Alexander, Denis R.

PY - 2002/1/18

Y1 - 2002/1/18

N2 - An investigation into the role of CD45 isoforms in T cell antigen receptor signal transduction was carried out by transfecting CD45-negative CD4+CD8+ HPB-ALL T cells with the CD45R0, CD45RBC, and CD45RABC isoforms. Fluorescence resonance energy transfer analysis showed that the CD45R0 isoform, but not the CD45RBC or CD45RABC isoforms, was found as homodimers and also preferentially associated with CD4 and CD8 at the cell-surface. A comparison was therefore made of T cell antigen receptor signaling between sub-clones expressing either CD45R0 or CD45RBC. Under basal conditions CD4-associated p56lck tyrosine kinase activity and cellular protein tyrosine phosphorylation levels were higher in the CD45R0+ than in the CD45RBC+ sub-clones. Upon CD3-CD4 ligation, TCR-ζ phosphorylation, ZAP-70 recruitment to the p21/p23 TCR-ζ phosphoisomers, ZAP-70 phosphorylation, as well as p56lck, c-Cbl and Slp-76 phosphorylation, were all markedly increased in CD45R0+ compared with CD45RBC+ cells. T cell antigen receptor (TCR) stimulation alone also promoted c-Cbl phosphorylation in CD45R0+ but not in CD45RBC+ cells. Our results are consistent with a model in which association of CD45R0 with CD4 generates a more active pool of CD4-associated p56lck kinase molecules. Upon CD3-CD4 co-ligation, the active p56lck increases the intensity of T cell antigen receptor signal transduction coupling by promoting TCR-ζ chain phosphorylation and ZAP-70 recruitment.

AB - An investigation into the role of CD45 isoforms in T cell antigen receptor signal transduction was carried out by transfecting CD45-negative CD4+CD8+ HPB-ALL T cells with the CD45R0, CD45RBC, and CD45RABC isoforms. Fluorescence resonance energy transfer analysis showed that the CD45R0 isoform, but not the CD45RBC or CD45RABC isoforms, was found as homodimers and also preferentially associated with CD4 and CD8 at the cell-surface. A comparison was therefore made of T cell antigen receptor signaling between sub-clones expressing either CD45R0 or CD45RBC. Under basal conditions CD4-associated p56lck tyrosine kinase activity and cellular protein tyrosine phosphorylation levels were higher in the CD45R0+ than in the CD45RBC+ sub-clones. Upon CD3-CD4 ligation, TCR-ζ phosphorylation, ZAP-70 recruitment to the p21/p23 TCR-ζ phosphoisomers, ZAP-70 phosphorylation, as well as p56lck, c-Cbl and Slp-76 phosphorylation, were all markedly increased in CD45R0+ compared with CD45RBC+ cells. T cell antigen receptor (TCR) stimulation alone also promoted c-Cbl phosphorylation in CD45R0+ but not in CD45RBC+ cells. Our results are consistent with a model in which association of CD45R0 with CD4 generates a more active pool of CD4-associated p56lck kinase molecules. Upon CD3-CD4 co-ligation, the active p56lck increases the intensity of T cell antigen receptor signal transduction coupling by promoting TCR-ζ chain phosphorylation and ZAP-70 recruitment.

UR - http://www.scopus.com/inward/record.url?scp=18544371851&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=18544371851&partnerID=8YFLogxK

U2 - 10.1074/jbc.M108386200

DO - 10.1074/jbc.M108386200

M3 - Article

C2 - 11694532

AN - SCOPUS:18544371851

VL - 277

SP - 1912

EP - 1918

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 3

ER -