A calcium requirement for jnk activation contributes to c-jun hyperexpression by P210 BCR-ABL

E. A. Williamson, M. J. Stewart, S. Litz-Jackson, G. S. Burgess, M. T. Rizzo, A. S. Kraft, H. Boswell

Research output: Contribution to journalArticle

Abstract

Hyperexpression of c-jun and c-fos mRNA's are cndpoints of the p21ras-dependent pathway of transformation by p210 BCR-ABL, the Philadelphia chromosome product. We studied the contribution of JNK, the cjun N-terminal kinase that augments, by phosphorylation, c-jun/AP-1-dependent transact!vation of c-jun transcription and elk-1 -dependent c-fos transcription, to explain c-jun and c-fos hyperexpression. By titration of cell extracts in a kinase assay involving GST-c-jun (5-89) as substrate, there was at least 6-fold greater activity of JNK within p210 BCR-ABL transformed cells compared to IL-3 treated parental cells. Corresponding to heightened JNK activity downstream of p210 BCR-ABL, there was a unique transactivating complex (AP-1 complex 2} consisting of c-jun and c-fos, binding to the proximal jun promoter (jun 1) TRE in gel mobility shifts. By contrast, the AP-1 contained within IL-3-dependent H7 parental cells was exclusively c-jun/ATF-2 heterodimer (complex 1). 5'-cjun mediated transcriptional activity in BCR-ABL transformed cells was totally dependent on binding of AP-1 by junl TRE, as demonstrated in reporter assays after site-directed mutagenesis of jun 1 TRE. Dose-dependent inhibition of JNK activity within BCR-ABL transformed cells resulted from treatment with cyclosporin A (CsA), an inhibitor of the Ca2+-calmodulin dependent enzyme, calcineurin (CN) [also called PP2B], or by treatment with W7, a calmodulin inhibitor. Dose-dependent inhibition of c-jun transcript expression on Northern blots followed JNK inhibition by CsA. Inhibition of c-jun mRNA by cyclosporin was a unique property of BCR-ABL transformed cells (absent in parental cells) and CsA treatment led to depletion of the unique c-jun/c-fos AP-1 complex (complex 2) binding to the 5' jun 1 site. Thus, BCR-ABL transformation requires JNK activity for c-jun and c-fos expression and a Ca2+-dependent JNK pathway unique to transformed cells is operating.

Original languageEnglish
Pages (from-to)1114
Number of pages1
JournalExperimental Hematology
Volume24
Issue number9
StatePublished - 1996

Fingerprint

Transcription Factor AP-1
Calcium
Cyclosporine
Interleukin-3
Calmodulin
Phosphotransferases
Philadelphia Chromosome
Messenger RNA
MAP Kinase Signaling System
Calcineurin
Site-Directed Mutagenesis
Cell Extracts
Northern Blotting
Gels
Phosphorylation
Enzymes

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

Williamson, E. A., Stewart, M. J., Litz-Jackson, S., Burgess, G. S., Rizzo, M. T., Kraft, A. S., & Boswell, H. (1996). A calcium requirement for jnk activation contributes to c-jun hyperexpression by P210 BCR-ABL. Experimental Hematology, 24(9), 1114.

A calcium requirement for jnk activation contributes to c-jun hyperexpression by P210 BCR-ABL. / Williamson, E. A.; Stewart, M. J.; Litz-Jackson, S.; Burgess, G. S.; Rizzo, M. T.; Kraft, A. S.; Boswell, H.

In: Experimental Hematology, Vol. 24, No. 9, 1996, p. 1114.

Research output: Contribution to journalArticle

Williamson, EA, Stewart, MJ, Litz-Jackson, S, Burgess, GS, Rizzo, MT, Kraft, AS & Boswell, H 1996, 'A calcium requirement for jnk activation contributes to c-jun hyperexpression by P210 BCR-ABL', Experimental Hematology, vol. 24, no. 9, pp. 1114.
Williamson EA, Stewart MJ, Litz-Jackson S, Burgess GS, Rizzo MT, Kraft AS et al. A calcium requirement for jnk activation contributes to c-jun hyperexpression by P210 BCR-ABL. Experimental Hematology. 1996;24(9):1114.
Williamson, E. A. ; Stewart, M. J. ; Litz-Jackson, S. ; Burgess, G. S. ; Rizzo, M. T. ; Kraft, A. S. ; Boswell, H. / A calcium requirement for jnk activation contributes to c-jun hyperexpression by P210 BCR-ABL. In: Experimental Hematology. 1996 ; Vol. 24, No. 9. pp. 1114.
@article{550c387efb564ba9a0fa22a7b71136f7,
title = "A calcium requirement for jnk activation contributes to c-jun hyperexpression by P210 BCR-ABL",
abstract = "Hyperexpression of c-jun and c-fos mRNA's are cndpoints of the p21ras-dependent pathway of transformation by p210 BCR-ABL, the Philadelphia chromosome product. We studied the contribution of JNK, the cjun N-terminal kinase that augments, by phosphorylation, c-jun/AP-1-dependent transact!vation of c-jun transcription and elk-1 -dependent c-fos transcription, to explain c-jun and c-fos hyperexpression. By titration of cell extracts in a kinase assay involving GST-c-jun (5-89) as substrate, there was at least 6-fold greater activity of JNK within p210 BCR-ABL transformed cells compared to IL-3 treated parental cells. Corresponding to heightened JNK activity downstream of p210 BCR-ABL, there was a unique transactivating complex (AP-1 complex 2} consisting of c-jun and c-fos, binding to the proximal jun promoter (jun 1) TRE in gel mobility shifts. By contrast, the AP-1 contained within IL-3-dependent H7 parental cells was exclusively c-jun/ATF-2 heterodimer (complex 1). 5'-cjun mediated transcriptional activity in BCR-ABL transformed cells was totally dependent on binding of AP-1 by junl TRE, as demonstrated in reporter assays after site-directed mutagenesis of jun 1 TRE. Dose-dependent inhibition of JNK activity within BCR-ABL transformed cells resulted from treatment with cyclosporin A (CsA), an inhibitor of the Ca2+-calmodulin dependent enzyme, calcineurin (CN) [also called PP2B], or by treatment with W7, a calmodulin inhibitor. Dose-dependent inhibition of c-jun transcript expression on Northern blots followed JNK inhibition by CsA. Inhibition of c-jun mRNA by cyclosporin was a unique property of BCR-ABL transformed cells (absent in parental cells) and CsA treatment led to depletion of the unique c-jun/c-fos AP-1 complex (complex 2) binding to the 5' jun 1 site. Thus, BCR-ABL transformation requires JNK activity for c-jun and c-fos expression and a Ca2+-dependent JNK pathway unique to transformed cells is operating.",
author = "Williamson, {E. A.} and Stewart, {M. J.} and S. Litz-Jackson and Burgess, {G. S.} and Rizzo, {M. T.} and Kraft, {A. S.} and H. Boswell",
year = "1996",
language = "English",
volume = "24",
pages = "1114",
journal = "Experimental Hematology",
issn = "0301-472X",
publisher = "Elsevier Inc.",
number = "9",

}

TY - JOUR

T1 - A calcium requirement for jnk activation contributes to c-jun hyperexpression by P210 BCR-ABL

AU - Williamson, E. A.

AU - Stewart, M. J.

AU - Litz-Jackson, S.

AU - Burgess, G. S.

AU - Rizzo, M. T.

AU - Kraft, A. S.

AU - Boswell, H.

PY - 1996

Y1 - 1996

N2 - Hyperexpression of c-jun and c-fos mRNA's are cndpoints of the p21ras-dependent pathway of transformation by p210 BCR-ABL, the Philadelphia chromosome product. We studied the contribution of JNK, the cjun N-terminal kinase that augments, by phosphorylation, c-jun/AP-1-dependent transact!vation of c-jun transcription and elk-1 -dependent c-fos transcription, to explain c-jun and c-fos hyperexpression. By titration of cell extracts in a kinase assay involving GST-c-jun (5-89) as substrate, there was at least 6-fold greater activity of JNK within p210 BCR-ABL transformed cells compared to IL-3 treated parental cells. Corresponding to heightened JNK activity downstream of p210 BCR-ABL, there was a unique transactivating complex (AP-1 complex 2} consisting of c-jun and c-fos, binding to the proximal jun promoter (jun 1) TRE in gel mobility shifts. By contrast, the AP-1 contained within IL-3-dependent H7 parental cells was exclusively c-jun/ATF-2 heterodimer (complex 1). 5'-cjun mediated transcriptional activity in BCR-ABL transformed cells was totally dependent on binding of AP-1 by junl TRE, as demonstrated in reporter assays after site-directed mutagenesis of jun 1 TRE. Dose-dependent inhibition of JNK activity within BCR-ABL transformed cells resulted from treatment with cyclosporin A (CsA), an inhibitor of the Ca2+-calmodulin dependent enzyme, calcineurin (CN) [also called PP2B], or by treatment with W7, a calmodulin inhibitor. Dose-dependent inhibition of c-jun transcript expression on Northern blots followed JNK inhibition by CsA. Inhibition of c-jun mRNA by cyclosporin was a unique property of BCR-ABL transformed cells (absent in parental cells) and CsA treatment led to depletion of the unique c-jun/c-fos AP-1 complex (complex 2) binding to the 5' jun 1 site. Thus, BCR-ABL transformation requires JNK activity for c-jun and c-fos expression and a Ca2+-dependent JNK pathway unique to transformed cells is operating.

AB - Hyperexpression of c-jun and c-fos mRNA's are cndpoints of the p21ras-dependent pathway of transformation by p210 BCR-ABL, the Philadelphia chromosome product. We studied the contribution of JNK, the cjun N-terminal kinase that augments, by phosphorylation, c-jun/AP-1-dependent transact!vation of c-jun transcription and elk-1 -dependent c-fos transcription, to explain c-jun and c-fos hyperexpression. By titration of cell extracts in a kinase assay involving GST-c-jun (5-89) as substrate, there was at least 6-fold greater activity of JNK within p210 BCR-ABL transformed cells compared to IL-3 treated parental cells. Corresponding to heightened JNK activity downstream of p210 BCR-ABL, there was a unique transactivating complex (AP-1 complex 2} consisting of c-jun and c-fos, binding to the proximal jun promoter (jun 1) TRE in gel mobility shifts. By contrast, the AP-1 contained within IL-3-dependent H7 parental cells was exclusively c-jun/ATF-2 heterodimer (complex 1). 5'-cjun mediated transcriptional activity in BCR-ABL transformed cells was totally dependent on binding of AP-1 by junl TRE, as demonstrated in reporter assays after site-directed mutagenesis of jun 1 TRE. Dose-dependent inhibition of JNK activity within BCR-ABL transformed cells resulted from treatment with cyclosporin A (CsA), an inhibitor of the Ca2+-calmodulin dependent enzyme, calcineurin (CN) [also called PP2B], or by treatment with W7, a calmodulin inhibitor. Dose-dependent inhibition of c-jun transcript expression on Northern blots followed JNK inhibition by CsA. Inhibition of c-jun mRNA by cyclosporin was a unique property of BCR-ABL transformed cells (absent in parental cells) and CsA treatment led to depletion of the unique c-jun/c-fos AP-1 complex (complex 2) binding to the 5' jun 1 site. Thus, BCR-ABL transformation requires JNK activity for c-jun and c-fos expression and a Ca2+-dependent JNK pathway unique to transformed cells is operating.

UR - http://www.scopus.com/inward/record.url?scp=33748607180&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748607180&partnerID=8YFLogxK

M3 - Article

VL - 24

SP - 1114

JO - Experimental Hematology

JF - Experimental Hematology

SN - 0301-472X

IS - 9

ER -