Prolactin stimulates activation of c-jun N-terminal kinase (JNK)

K. L. Schwertfeger, S. Hunter, L. E. Heasley, V. Levresse, R. P. Leon, J. DeGregori, S. M. Anderson

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

In recent years the mitogen-activated protein (MAP) kinase family has expanded to include both c-jun N-terminal kinases (JNKs), and the p38/HOG1 family in addition to the extracellular regulated kinase (ERK) family. These kinases are activated by a variety of growth factors, as well as extra- and intracellular insults such as osmotic stress, UV light, and chemotherapeutic agents. Stimulation of the PRL-dependent Nb2 cell line with PRL results in the rapid activation of JNK as determined by the glutathione-S-transferase (GST)-jun kinase assay. Activation was maximal 30 min after stimulation with 50 nM rat PRL (rPRL) and decreased after that time. Dose response studies indicated that concentrations as low as 10 nM rPRL resulted in maximal activation. The interleukin-3 (IL-3)-dependent myeloid progenitor cell line 32Dcl3 was transfected with the long, Nb2, and short forms of the rat PRL receptor (rPRLR), as well as the long form of the human PRLR (hPRLR). The long and Nb2 forms of the PRLR were able to stimulate activation of JNK; however, the short form of the rPRLR was not. This corresponds with the inability of the short form of the rPRLR to stimulate proliferation of 32Dcl3 cells. Activation of JNK in 32Dcl3 cells expressing the long form of the hPRLR was maximal at 30 min after stimulation with 100 nM ovine PRL (oPRL) and declined after that time. Dose response studies indicated that activation of JNK was maximal after 30 min at a concentration of 10 nM, and the amount of activated JNK declined at the highest concentration of oPRL, 100 nM. Immunoblot analysis with an antibody that recognizes the activated (phosphorylated) forms of JNK1 and JNK2 indicated that both JNK1 and JNK2 isoforms were activated in 32D/hPRLR cells stimulated with oPRL. A recombinant human adenovirus expressing a kinase-inactive mutant of JNK1 (APF mutant) was used to determine the biological effect of blocking JNK activity in Nb2 cells. Expression of the JNK1-APF mutant inhibited cellular proliferation and induced DNA fragmentation typical of cells undergoing apoptosis. These data suggest that activation of JNKs may be important in mitogenic signaling and/or suppression of apoptosis in Nb2 cells.

Original languageEnglish (US)
Pages (from-to)1592-1602
Number of pages11
JournalMolecular Endocrinology
Volume14
Issue number10
DOIs
StatePublished - Jan 1 2000

Fingerprint

JNK Mitogen-Activated Protein Kinases
Prolactin
Prolactin Receptors
Phosphotransferases
Sheep
Cell Proliferation
Apoptosis
Myeloid Progenitor Cells
Human Adenoviruses
Cell Line
Interleukin-3
Osmotic Pressure
DNA Fragmentation
Ultraviolet Rays
Mitogen-Activated Protein Kinases
Glutathione Transferase
Intercellular Signaling Peptides and Proteins
Protein Isoforms
Antibodies

Cite this

Schwertfeger, K. L., Hunter, S., Heasley, L. E., Levresse, V., Leon, R. P., DeGregori, J., & Anderson, S. M. (2000). Prolactin stimulates activation of c-jun N-terminal kinase (JNK). Molecular Endocrinology, 14(10), 1592-1602. https://doi.org/10.1210/mend.14.10.0536

Prolactin stimulates activation of c-jun N-terminal kinase (JNK). / Schwertfeger, K. L.; Hunter, S.; Heasley, L. E.; Levresse, V.; Leon, R. P.; DeGregori, J.; Anderson, S. M.

In: Molecular Endocrinology, Vol. 14, No. 10, 01.01.2000, p. 1592-1602.

Research output: Contribution to journalArticle

Schwertfeger, KL, Hunter, S, Heasley, LE, Levresse, V, Leon, RP, DeGregori, J & Anderson, SM 2000, 'Prolactin stimulates activation of c-jun N-terminal kinase (JNK)', Molecular Endocrinology, vol. 14, no. 10, pp. 1592-1602. https://doi.org/10.1210/mend.14.10.0536
Schwertfeger KL, Hunter S, Heasley LE, Levresse V, Leon RP, DeGregori J et al. Prolactin stimulates activation of c-jun N-terminal kinase (JNK). Molecular Endocrinology. 2000 Jan 1;14(10):1592-1602. https://doi.org/10.1210/mend.14.10.0536
Schwertfeger, K. L. ; Hunter, S. ; Heasley, L. E. ; Levresse, V. ; Leon, R. P. ; DeGregori, J. ; Anderson, S. M. / Prolactin stimulates activation of c-jun N-terminal kinase (JNK). In: Molecular Endocrinology. 2000 ; Vol. 14, No. 10. pp. 1592-1602.
@article{03f984ed181144c29287c1cdb393fd43,
title = "Prolactin stimulates activation of c-jun N-terminal kinase (JNK)",
abstract = "In recent years the mitogen-activated protein (MAP) kinase family has expanded to include both c-jun N-terminal kinases (JNKs), and the p38/HOG1 family in addition to the extracellular regulated kinase (ERK) family. These kinases are activated by a variety of growth factors, as well as extra- and intracellular insults such as osmotic stress, UV light, and chemotherapeutic agents. Stimulation of the PRL-dependent Nb2 cell line with PRL results in the rapid activation of JNK as determined by the glutathione-S-transferase (GST)-jun kinase assay. Activation was maximal 30 min after stimulation with 50 nM rat PRL (rPRL) and decreased after that time. Dose response studies indicated that concentrations as low as 10 nM rPRL resulted in maximal activation. The interleukin-3 (IL-3)-dependent myeloid progenitor cell line 32Dcl3 was transfected with the long, Nb2, and short forms of the rat PRL receptor (rPRLR), as well as the long form of the human PRLR (hPRLR). The long and Nb2 forms of the PRLR were able to stimulate activation of JNK; however, the short form of the rPRLR was not. This corresponds with the inability of the short form of the rPRLR to stimulate proliferation of 32Dcl3 cells. Activation of JNK in 32Dcl3 cells expressing the long form of the hPRLR was maximal at 30 min after stimulation with 100 nM ovine PRL (oPRL) and declined after that time. Dose response studies indicated that activation of JNK was maximal after 30 min at a concentration of 10 nM, and the amount of activated JNK declined at the highest concentration of oPRL, 100 nM. Immunoblot analysis with an antibody that recognizes the activated (phosphorylated) forms of JNK1 and JNK2 indicated that both JNK1 and JNK2 isoforms were activated in 32D/hPRLR cells stimulated with oPRL. A recombinant human adenovirus expressing a kinase-inactive mutant of JNK1 (APF mutant) was used to determine the biological effect of blocking JNK activity in Nb2 cells. Expression of the JNK1-APF mutant inhibited cellular proliferation and induced DNA fragmentation typical of cells undergoing apoptosis. These data suggest that activation of JNKs may be important in mitogenic signaling and/or suppression of apoptosis in Nb2 cells.",
author = "Schwertfeger, {K. L.} and S. Hunter and Heasley, {L. E.} and V. Levresse and Leon, {R. P.} and J. DeGregori and Anderson, {S. M.}",
year = "2000",
month = "1",
day = "1",
doi = "10.1210/mend.14.10.0536",
language = "English (US)",
volume = "14",
pages = "1592--1602",
journal = "Molecular Endocrinology",
issn = "0888-8809",
publisher = "The Endocrine Society",
number = "10",

}

TY - JOUR

T1 - Prolactin stimulates activation of c-jun N-terminal kinase (JNK)

AU - Schwertfeger, K. L.

AU - Hunter, S.

AU - Heasley, L. E.

AU - Levresse, V.

AU - Leon, R. P.

AU - DeGregori, J.

AU - Anderson, S. M.

PY - 2000/1/1

Y1 - 2000/1/1

N2 - In recent years the mitogen-activated protein (MAP) kinase family has expanded to include both c-jun N-terminal kinases (JNKs), and the p38/HOG1 family in addition to the extracellular regulated kinase (ERK) family. These kinases are activated by a variety of growth factors, as well as extra- and intracellular insults such as osmotic stress, UV light, and chemotherapeutic agents. Stimulation of the PRL-dependent Nb2 cell line with PRL results in the rapid activation of JNK as determined by the glutathione-S-transferase (GST)-jun kinase assay. Activation was maximal 30 min after stimulation with 50 nM rat PRL (rPRL) and decreased after that time. Dose response studies indicated that concentrations as low as 10 nM rPRL resulted in maximal activation. The interleukin-3 (IL-3)-dependent myeloid progenitor cell line 32Dcl3 was transfected with the long, Nb2, and short forms of the rat PRL receptor (rPRLR), as well as the long form of the human PRLR (hPRLR). The long and Nb2 forms of the PRLR were able to stimulate activation of JNK; however, the short form of the rPRLR was not. This corresponds with the inability of the short form of the rPRLR to stimulate proliferation of 32Dcl3 cells. Activation of JNK in 32Dcl3 cells expressing the long form of the hPRLR was maximal at 30 min after stimulation with 100 nM ovine PRL (oPRL) and declined after that time. Dose response studies indicated that activation of JNK was maximal after 30 min at a concentration of 10 nM, and the amount of activated JNK declined at the highest concentration of oPRL, 100 nM. Immunoblot analysis with an antibody that recognizes the activated (phosphorylated) forms of JNK1 and JNK2 indicated that both JNK1 and JNK2 isoforms were activated in 32D/hPRLR cells stimulated with oPRL. A recombinant human adenovirus expressing a kinase-inactive mutant of JNK1 (APF mutant) was used to determine the biological effect of blocking JNK activity in Nb2 cells. Expression of the JNK1-APF mutant inhibited cellular proliferation and induced DNA fragmentation typical of cells undergoing apoptosis. These data suggest that activation of JNKs may be important in mitogenic signaling and/or suppression of apoptosis in Nb2 cells.

AB - In recent years the mitogen-activated protein (MAP) kinase family has expanded to include both c-jun N-terminal kinases (JNKs), and the p38/HOG1 family in addition to the extracellular regulated kinase (ERK) family. These kinases are activated by a variety of growth factors, as well as extra- and intracellular insults such as osmotic stress, UV light, and chemotherapeutic agents. Stimulation of the PRL-dependent Nb2 cell line with PRL results in the rapid activation of JNK as determined by the glutathione-S-transferase (GST)-jun kinase assay. Activation was maximal 30 min after stimulation with 50 nM rat PRL (rPRL) and decreased after that time. Dose response studies indicated that concentrations as low as 10 nM rPRL resulted in maximal activation. The interleukin-3 (IL-3)-dependent myeloid progenitor cell line 32Dcl3 was transfected with the long, Nb2, and short forms of the rat PRL receptor (rPRLR), as well as the long form of the human PRLR (hPRLR). The long and Nb2 forms of the PRLR were able to stimulate activation of JNK; however, the short form of the rPRLR was not. This corresponds with the inability of the short form of the rPRLR to stimulate proliferation of 32Dcl3 cells. Activation of JNK in 32Dcl3 cells expressing the long form of the hPRLR was maximal at 30 min after stimulation with 100 nM ovine PRL (oPRL) and declined after that time. Dose response studies indicated that activation of JNK was maximal after 30 min at a concentration of 10 nM, and the amount of activated JNK declined at the highest concentration of oPRL, 100 nM. Immunoblot analysis with an antibody that recognizes the activated (phosphorylated) forms of JNK1 and JNK2 indicated that both JNK1 and JNK2 isoforms were activated in 32D/hPRLR cells stimulated with oPRL. A recombinant human adenovirus expressing a kinase-inactive mutant of JNK1 (APF mutant) was used to determine the biological effect of blocking JNK activity in Nb2 cells. Expression of the JNK1-APF mutant inhibited cellular proliferation and induced DNA fragmentation typical of cells undergoing apoptosis. These data suggest that activation of JNKs may be important in mitogenic signaling and/or suppression of apoptosis in Nb2 cells.

UR - http://www.scopus.com/inward/record.url?scp=0033777436&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033777436&partnerID=8YFLogxK

U2 - 10.1210/mend.14.10.0536

DO - 10.1210/mend.14.10.0536

M3 - Article

C2 - 11043575

AN - SCOPUS:0033777436

VL - 14

SP - 1592

EP - 1602

JO - Molecular Endocrinology

JF - Molecular Endocrinology

SN - 0888-8809

IS - 10

ER -