CD2 signalling induces phosphorylation of CREB in primary lymphocytes

D. J. Guyot, G. C. Newbound, Michael Dale Lairmore

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Promoter sequences responsive to cyclic AMP (cAMP) are found in a number of cellular genes, and bind transcription factors of the cAMP response element binding protein (CREB)/activating transcription factor-1 (ATF-1) family. We have used a human T-lymphotropic virus type 1 (HTLV-1) model of cAMP response element (CRE) transcription to investigate the influence of lymphocyte activation on transcription from homologous regions in the viral promoter. We previously demonstrated increased HTLV-1 transcription following CD2 but not CD3 receptor cross-linking. We hypothesized that this increased viral transcription was mediated, in part, through the phosphorylation of CREB. Therefore, we investigated CD2 and CD3 receptor-mediated signalling in primary human peripheral blood mononuclear cells (PBMC). CD2, but not CD3, cross-linking increased cAMP detected by competitive enzyme-linked immunosorbent assay (ELISA) approximately fourfold. CD2 cross-linking concurrently increased phosphorylation of CREB detected by immunoblot assay eightfold. Consistent with post-translational regulation, no change in total level of CREB protein was observed. Phosphorylation of CREB occurred through a herbimycin A and Rp-cAMP-sensitive pathway, suggesting phosphorylation required antecedent activation of both protein tyrosine kinases (PTK) and protein kinase A (PKA). Both CD2 and CD3 cross-linking increased binding of nuclear proteins to a radiolabelled CRE oligonucleotide probe in electrophoretic mobility shift assays suggesting that lymphocyte activation enhances binding independently of phosphorylation of CREB at serine 133. These data indicate specific modulation of the CREB/ATF-1 family of transcription factors by the CD2 signalling pathway and suggest CD2 receptor modulation of CRE-mediated transcription following ligand engagement (e.g. cell-to-cell contact).

Original languageEnglish (US)
Pages (from-to)544-552
Number of pages9
JournalImmunology
Volume95
Issue number4
DOIs
StatePublished - 1998
Externally publishedYes

Fingerprint

Cyclic AMP Response Element-Binding Protein
Phosphorylation
Lymphocytes
Cyclic AMP
Activating Transcription Factor 1
Response Elements
Human T-lymphotropic virus 1
Lymphocyte Activation
Transcription Factors
Cyclic AMP Receptors
Oligonucleotide Probes
Electrophoretic Mobility Shift Assay
Nuclear Proteins
Cyclic AMP-Dependent Protein Kinases
Protein-Tyrosine Kinases
Serine
Blood Cells
Carrier Proteins
Enzyme-Linked Immunosorbent Assay
Ligands

ASJC Scopus subject areas

  • Immunology

Cite this

CD2 signalling induces phosphorylation of CREB in primary lymphocytes. / Guyot, D. J.; Newbound, G. C.; Lairmore, Michael Dale.

In: Immunology, Vol. 95, No. 4, 1998, p. 544-552.

Research output: Contribution to journalArticle

Guyot, D. J. ; Newbound, G. C. ; Lairmore, Michael Dale. / CD2 signalling induces phosphorylation of CREB in primary lymphocytes. In: Immunology. 1998 ; Vol. 95, No. 4. pp. 544-552.
@article{d8bd463aabd04e49bf426ecc0ad459f8,
title = "CD2 signalling induces phosphorylation of CREB in primary lymphocytes",
abstract = "Promoter sequences responsive to cyclic AMP (cAMP) are found in a number of cellular genes, and bind transcription factors of the cAMP response element binding protein (CREB)/activating transcription factor-1 (ATF-1) family. We have used a human T-lymphotropic virus type 1 (HTLV-1) model of cAMP response element (CRE) transcription to investigate the influence of lymphocyte activation on transcription from homologous regions in the viral promoter. We previously demonstrated increased HTLV-1 transcription following CD2 but not CD3 receptor cross-linking. We hypothesized that this increased viral transcription was mediated, in part, through the phosphorylation of CREB. Therefore, we investigated CD2 and CD3 receptor-mediated signalling in primary human peripheral blood mononuclear cells (PBMC). CD2, but not CD3, cross-linking increased cAMP detected by competitive enzyme-linked immunosorbent assay (ELISA) approximately fourfold. CD2 cross-linking concurrently increased phosphorylation of CREB detected by immunoblot assay eightfold. Consistent with post-translational regulation, no change in total level of CREB protein was observed. Phosphorylation of CREB occurred through a herbimycin A and Rp-cAMP-sensitive pathway, suggesting phosphorylation required antecedent activation of both protein tyrosine kinases (PTK) and protein kinase A (PKA). Both CD2 and CD3 cross-linking increased binding of nuclear proteins to a radiolabelled CRE oligonucleotide probe in electrophoretic mobility shift assays suggesting that lymphocyte activation enhances binding independently of phosphorylation of CREB at serine 133. These data indicate specific modulation of the CREB/ATF-1 family of transcription factors by the CD2 signalling pathway and suggest CD2 receptor modulation of CRE-mediated transcription following ligand engagement (e.g. cell-to-cell contact).",
author = "Guyot, {D. J.} and Newbound, {G. C.} and Lairmore, {Michael Dale}",
year = "1998",
doi = "10.1046/j.1365-2567.1998.00632.x",
language = "English (US)",
volume = "95",
pages = "544--552",
journal = "Immunology",
issn = "0019-2805",
publisher = "Wiley-Blackwell",
number = "4",

}

TY - JOUR

T1 - CD2 signalling induces phosphorylation of CREB in primary lymphocytes

AU - Guyot, D. J.

AU - Newbound, G. C.

AU - Lairmore, Michael Dale

PY - 1998

Y1 - 1998

N2 - Promoter sequences responsive to cyclic AMP (cAMP) are found in a number of cellular genes, and bind transcription factors of the cAMP response element binding protein (CREB)/activating transcription factor-1 (ATF-1) family. We have used a human T-lymphotropic virus type 1 (HTLV-1) model of cAMP response element (CRE) transcription to investigate the influence of lymphocyte activation on transcription from homologous regions in the viral promoter. We previously demonstrated increased HTLV-1 transcription following CD2 but not CD3 receptor cross-linking. We hypothesized that this increased viral transcription was mediated, in part, through the phosphorylation of CREB. Therefore, we investigated CD2 and CD3 receptor-mediated signalling in primary human peripheral blood mononuclear cells (PBMC). CD2, but not CD3, cross-linking increased cAMP detected by competitive enzyme-linked immunosorbent assay (ELISA) approximately fourfold. CD2 cross-linking concurrently increased phosphorylation of CREB detected by immunoblot assay eightfold. Consistent with post-translational regulation, no change in total level of CREB protein was observed. Phosphorylation of CREB occurred through a herbimycin A and Rp-cAMP-sensitive pathway, suggesting phosphorylation required antecedent activation of both protein tyrosine kinases (PTK) and protein kinase A (PKA). Both CD2 and CD3 cross-linking increased binding of nuclear proteins to a radiolabelled CRE oligonucleotide probe in electrophoretic mobility shift assays suggesting that lymphocyte activation enhances binding independently of phosphorylation of CREB at serine 133. These data indicate specific modulation of the CREB/ATF-1 family of transcription factors by the CD2 signalling pathway and suggest CD2 receptor modulation of CRE-mediated transcription following ligand engagement (e.g. cell-to-cell contact).

AB - Promoter sequences responsive to cyclic AMP (cAMP) are found in a number of cellular genes, and bind transcription factors of the cAMP response element binding protein (CREB)/activating transcription factor-1 (ATF-1) family. We have used a human T-lymphotropic virus type 1 (HTLV-1) model of cAMP response element (CRE) transcription to investigate the influence of lymphocyte activation on transcription from homologous regions in the viral promoter. We previously demonstrated increased HTLV-1 transcription following CD2 but not CD3 receptor cross-linking. We hypothesized that this increased viral transcription was mediated, in part, through the phosphorylation of CREB. Therefore, we investigated CD2 and CD3 receptor-mediated signalling in primary human peripheral blood mononuclear cells (PBMC). CD2, but not CD3, cross-linking increased cAMP detected by competitive enzyme-linked immunosorbent assay (ELISA) approximately fourfold. CD2 cross-linking concurrently increased phosphorylation of CREB detected by immunoblot assay eightfold. Consistent with post-translational regulation, no change in total level of CREB protein was observed. Phosphorylation of CREB occurred through a herbimycin A and Rp-cAMP-sensitive pathway, suggesting phosphorylation required antecedent activation of both protein tyrosine kinases (PTK) and protein kinase A (PKA). Both CD2 and CD3 cross-linking increased binding of nuclear proteins to a radiolabelled CRE oligonucleotide probe in electrophoretic mobility shift assays suggesting that lymphocyte activation enhances binding independently of phosphorylation of CREB at serine 133. These data indicate specific modulation of the CREB/ATF-1 family of transcription factors by the CD2 signalling pathway and suggest CD2 receptor modulation of CRE-mediated transcription following ligand engagement (e.g. cell-to-cell contact).

UR - http://www.scopus.com/inward/record.url?scp=0031797927&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031797927&partnerID=8YFLogxK

U2 - 10.1046/j.1365-2567.1998.00632.x

DO - 10.1046/j.1365-2567.1998.00632.x

M3 - Article

C2 - 9893043

AN - SCOPUS:0031797927

VL - 95

SP - 544

EP - 552

JO - Immunology

JF - Immunology

SN - 0019-2805

IS - 4

ER -