Activation of the aryl hydrocarbon receptor by the widely used Src family kinase inhibitor 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine (PP2)

Katrin Frauenstein, Julia Tigges, Anatoly A. Soshilov, Sarah Kado, Nadeshda Raab, Ellen Fritsche, Judith Haendeler, Michael S. Denison, Christoph F A Vogel, Thomas Haarmann-Stemmann

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Small molecular weight protein kinase inhibitors are frequently used tools to unravel the complex network of cellular signal transduction under certain physiological and pathophysiological conditions. 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine (PP2) is a widely used compound to block the activity of Src family kinases, the major group of non-receptor tyrosine kinases, which trigger multiple cellular signaling pathways. Here, we show that PP2 induces cytochrome P450 1A1 mRNA expression and enzyme activity in a dose-dependent manner in human HepG2 hepatoma cells and NCTC 2544 keratinocytes. By means of reporter gene assays, RNA interference, electrophoretic mobility shift assay, and competitive ligand-binding assay, we further demonstrate that PP2 is a ligand for the aryl hydrocarbon receptor (AHR), an intracellular chemosensor that regulates xenobiotic metabolism, environmental stress responses, and immune functions. Upon ligand-dependent activation, the AHR translocates into the nucleus and dimerizes with the AHR nuclear translocator (ARNT) to modulate the expression of its target genes. In addition, AHR activation is frequently accompanied by an activation of the tyrosine kinase c-Src, resulting in stimulation of cell-surface receptors and downstream signal transduction. As PP2 activates the AHR/ARNT pathway by simultaneously blocking c-Src-mediated alternative signaling routes, this compound may be a suitable tool to study the contribution of the different AHR-dependent signaling pathways to biological processes and adverse outcomes. On the other hand, the unexpected property of PP2 to stimulate AHR/ARNT signaling should be carefully taken into account in future investigations in order to avoid a false interpretation of experimental results and molecular interrelations.

Original languageEnglish (US)
Pages (from-to)1329-1336
Number of pages8
JournalArchives of Toxicology
Volume89
Issue number8
DOIs
StatePublished - Aug 1 2014

Fingerprint

pyrazolo(3,4-d)pyrimidine
Aryl Hydrocarbon Receptors
Aryl Hydrocarbon Receptor Nuclear Translocator
src-Family Kinases
Chemical activation
Ligands
Assays
Signal Transduction
Signal transduction
Biological Phenomena
Competitive Binding
Hep G2 Cells
Cell Surface Receptors
Electrophoretic Mobility Shift Assay
Xenobiotics
Protein Kinase Inhibitors
RNA Interference
Genes
Keratinocytes
Reporter Genes

Keywords

  • Aryl hydrocarbon receptor
  • CYP1A1
  • Off-target effect
  • PP2
  • Src family kinase

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Activation of the aryl hydrocarbon receptor by the widely used Src family kinase inhibitor 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine (PP2). / Frauenstein, Katrin; Tigges, Julia; Soshilov, Anatoly A.; Kado, Sarah; Raab, Nadeshda; Fritsche, Ellen; Haendeler, Judith; Denison, Michael S.; Vogel, Christoph F A; Haarmann-Stemmann, Thomas.

In: Archives of Toxicology, Vol. 89, No. 8, 01.08.2014, p. 1329-1336.

Research output: Contribution to journalArticle

Frauenstein, K, Tigges, J, Soshilov, AA, Kado, S, Raab, N, Fritsche, E, Haendeler, J, Denison, MS, Vogel, CFA & Haarmann-Stemmann, T 2014, 'Activation of the aryl hydrocarbon receptor by the widely used Src family kinase inhibitor 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine (PP2)', Archives of Toxicology, vol. 89, no. 8, pp. 1329-1336. https://doi.org/10.1007/s00204-014-1321-8
Frauenstein, Katrin ; Tigges, Julia ; Soshilov, Anatoly A. ; Kado, Sarah ; Raab, Nadeshda ; Fritsche, Ellen ; Haendeler, Judith ; Denison, Michael S. ; Vogel, Christoph F A ; Haarmann-Stemmann, Thomas. / Activation of the aryl hydrocarbon receptor by the widely used Src family kinase inhibitor 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine (PP2). In: Archives of Toxicology. 2014 ; Vol. 89, No. 8. pp. 1329-1336.
@article{4d03776d5d284c7c9d670d1d0b2f93c0,
title = "Activation of the aryl hydrocarbon receptor by the widely used Src family kinase inhibitor 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine (PP2)",
abstract = "Small molecular weight protein kinase inhibitors are frequently used tools to unravel the complex network of cellular signal transduction under certain physiological and pathophysiological conditions. 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine (PP2) is a widely used compound to block the activity of Src family kinases, the major group of non-receptor tyrosine kinases, which trigger multiple cellular signaling pathways. Here, we show that PP2 induces cytochrome P450 1A1 mRNA expression and enzyme activity in a dose-dependent manner in human HepG2 hepatoma cells and NCTC 2544 keratinocytes. By means of reporter gene assays, RNA interference, electrophoretic mobility shift assay, and competitive ligand-binding assay, we further demonstrate that PP2 is a ligand for the aryl hydrocarbon receptor (AHR), an intracellular chemosensor that regulates xenobiotic metabolism, environmental stress responses, and immune functions. Upon ligand-dependent activation, the AHR translocates into the nucleus and dimerizes with the AHR nuclear translocator (ARNT) to modulate the expression of its target genes. In addition, AHR activation is frequently accompanied by an activation of the tyrosine kinase c-Src, resulting in stimulation of cell-surface receptors and downstream signal transduction. As PP2 activates the AHR/ARNT pathway by simultaneously blocking c-Src-mediated alternative signaling routes, this compound may be a suitable tool to study the contribution of the different AHR-dependent signaling pathways to biological processes and adverse outcomes. On the other hand, the unexpected property of PP2 to stimulate AHR/ARNT signaling should be carefully taken into account in future investigations in order to avoid a false interpretation of experimental results and molecular interrelations.",
keywords = "Aryl hydrocarbon receptor, CYP1A1, Off-target effect, PP2, Src family kinase",
author = "Katrin Frauenstein and Julia Tigges and Soshilov, {Anatoly A.} and Sarah Kado and Nadeshda Raab and Ellen Fritsche and Judith Haendeler and Denison, {Michael S.} and Vogel, {Christoph F A} and Thomas Haarmann-Stemmann",
year = "2014",
month = "8",
day = "1",
doi = "10.1007/s00204-014-1321-8",
language = "English (US)",
volume = "89",
pages = "1329--1336",
journal = "Archiv fur Toxikologie",
issn = "0003-9446",
publisher = "Springer Verlag",
number = "8",

}

TY - JOUR

T1 - Activation of the aryl hydrocarbon receptor by the widely used Src family kinase inhibitor 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine (PP2)

AU - Frauenstein, Katrin

AU - Tigges, Julia

AU - Soshilov, Anatoly A.

AU - Kado, Sarah

AU - Raab, Nadeshda

AU - Fritsche, Ellen

AU - Haendeler, Judith

AU - Denison, Michael S.

AU - Vogel, Christoph F A

AU - Haarmann-Stemmann, Thomas

PY - 2014/8/1

Y1 - 2014/8/1

N2 - Small molecular weight protein kinase inhibitors are frequently used tools to unravel the complex network of cellular signal transduction under certain physiological and pathophysiological conditions. 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine (PP2) is a widely used compound to block the activity of Src family kinases, the major group of non-receptor tyrosine kinases, which trigger multiple cellular signaling pathways. Here, we show that PP2 induces cytochrome P450 1A1 mRNA expression and enzyme activity in a dose-dependent manner in human HepG2 hepatoma cells and NCTC 2544 keratinocytes. By means of reporter gene assays, RNA interference, electrophoretic mobility shift assay, and competitive ligand-binding assay, we further demonstrate that PP2 is a ligand for the aryl hydrocarbon receptor (AHR), an intracellular chemosensor that regulates xenobiotic metabolism, environmental stress responses, and immune functions. Upon ligand-dependent activation, the AHR translocates into the nucleus and dimerizes with the AHR nuclear translocator (ARNT) to modulate the expression of its target genes. In addition, AHR activation is frequently accompanied by an activation of the tyrosine kinase c-Src, resulting in stimulation of cell-surface receptors and downstream signal transduction. As PP2 activates the AHR/ARNT pathway by simultaneously blocking c-Src-mediated alternative signaling routes, this compound may be a suitable tool to study the contribution of the different AHR-dependent signaling pathways to biological processes and adverse outcomes. On the other hand, the unexpected property of PP2 to stimulate AHR/ARNT signaling should be carefully taken into account in future investigations in order to avoid a false interpretation of experimental results and molecular interrelations.

AB - Small molecular weight protein kinase inhibitors are frequently used tools to unravel the complex network of cellular signal transduction under certain physiological and pathophysiological conditions. 4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine (PP2) is a widely used compound to block the activity of Src family kinases, the major group of non-receptor tyrosine kinases, which trigger multiple cellular signaling pathways. Here, we show that PP2 induces cytochrome P450 1A1 mRNA expression and enzyme activity in a dose-dependent manner in human HepG2 hepatoma cells and NCTC 2544 keratinocytes. By means of reporter gene assays, RNA interference, electrophoretic mobility shift assay, and competitive ligand-binding assay, we further demonstrate that PP2 is a ligand for the aryl hydrocarbon receptor (AHR), an intracellular chemosensor that regulates xenobiotic metabolism, environmental stress responses, and immune functions. Upon ligand-dependent activation, the AHR translocates into the nucleus and dimerizes with the AHR nuclear translocator (ARNT) to modulate the expression of its target genes. In addition, AHR activation is frequently accompanied by an activation of the tyrosine kinase c-Src, resulting in stimulation of cell-surface receptors and downstream signal transduction. As PP2 activates the AHR/ARNT pathway by simultaneously blocking c-Src-mediated alternative signaling routes, this compound may be a suitable tool to study the contribution of the different AHR-dependent signaling pathways to biological processes and adverse outcomes. On the other hand, the unexpected property of PP2 to stimulate AHR/ARNT signaling should be carefully taken into account in future investigations in order to avoid a false interpretation of experimental results and molecular interrelations.

KW - Aryl hydrocarbon receptor

KW - CYP1A1

KW - Off-target effect

KW - PP2

KW - Src family kinase

UR - http://www.scopus.com/inward/record.url?scp=84937978414&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84937978414&partnerID=8YFLogxK

U2 - 10.1007/s00204-014-1321-8

DO - 10.1007/s00204-014-1321-8

M3 - Article

C2 - 25082669

AN - SCOPUS:84937978414

VL - 89

SP - 1329

EP - 1336

JO - Archiv fur Toxikologie

JF - Archiv fur Toxikologie

SN - 0003-9446

IS - 8

ER -