Proteasomes regulate erythropoietin receptor and signal transducer and activator of transcription 5 (STAT5) activation

Possible involvement of the ubiquitinated Cis protein

Frédérique Verdier, Stany Chrétien, Odile Muller, Paule Varlet, Akihiko Yoshimura, Sylvie Gisselbrecht, Catherine Lacombe, Patrick Mayeux

Research output: Contribution to journalArticle

150 Citations (Scopus)

Abstract

Cis is an Src homology 2 domain-containing protein, which binds to the erythropoietin receptor and decreases erythropoietin-stimulated cell proliferation. We show that Cis associates with the second tyrosine residue of the intracellular domain of the erythropoietin receptor (Tyr401). Two forms of Cis with molecular masses of 32 and 37 kDa were detected, and we demonstrate that the 37-kDa protein resulted from post-translational modifications of the 32-kDa form. Anti-ubiquitin antibodies recognized the 37-kDa form of Cis and the proteasome inhibitors N-acetyl-leucyl-leucyl-nor- leucinal and lactacystin inhibited its degradation, showing that the 37-kDa form of Cis is a ubiquitinated protein, which seems to be rapidly degraded by the proteasome. In erythropoietin-stimulated UT-7 cells, the activation of the erythropoietin receptor and signal transducer and activator of transcription 5 (STAT5) was transient and returned to basal levels after 30- 60 min of erythropoietin stimulation. In contrast, these proteins remained strongly phosphorylated, and STAT5 remained activated for at least 120 min in the presence of proteasome inhibitors. These experiments demonstrate that the proteasomes are involved in the down-regulation of the erythropoietin receptor activation signals. Because the proteasome inhibitors induced the accumulation of both the ubiquitinated form of Cis and the Cis- erythropoietin receptor complexes, our results suggest that the ubiquitinated form of Cis could be involved in the proteasome-mediated inactivation of the erythropoietin receptor.

Original languageEnglish
Pages (from-to)28185-28190
Number of pages6
JournalJournal of Biological Chemistry
Volume273
Issue number43
DOIs
Publication statusPublished - 1998 Oct 23
Externally publishedYes

Fingerprint

STAT5 Transcription Factor
Ubiquitinated Proteins
Erythropoietin Receptors
Proteasome Endopeptidase Complex
Transcriptional Activation
Chemical activation
Proteasome Inhibitors
Erythropoietin
Proteins
src Homology Domains
Cell proliferation
Molecular mass
Post Translational Protein Processing
Ubiquitin
Tyrosine
Anti-Idiotypic Antibodies
Down-Regulation
Cell Proliferation
Degradation
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Proteasomes regulate erythropoietin receptor and signal transducer and activator of transcription 5 (STAT5) activation : Possible involvement of the ubiquitinated Cis protein. / Verdier, Frédérique; Chrétien, Stany; Muller, Odile; Varlet, Paule; Yoshimura, Akihiko; Gisselbrecht, Sylvie; Lacombe, Catherine; Mayeux, Patrick.

In: Journal of Biological Chemistry, Vol. 273, No. 43, 23.10.1998, p. 28185-28190.

Research output: Contribution to journalArticle

Verdier, Frédérique ; Chrétien, Stany ; Muller, Odile ; Varlet, Paule ; Yoshimura, Akihiko ; Gisselbrecht, Sylvie ; Lacombe, Catherine ; Mayeux, Patrick. / Proteasomes regulate erythropoietin receptor and signal transducer and activator of transcription 5 (STAT5) activation : Possible involvement of the ubiquitinated Cis protein. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 43. pp. 28185-28190.
@article{5a525225c5844bb7a6c8c12e006ba26b,
title = "Proteasomes regulate erythropoietin receptor and signal transducer and activator of transcription 5 (STAT5) activation: Possible involvement of the ubiquitinated Cis protein",
abstract = "Cis is an Src homology 2 domain-containing protein, which binds to the erythropoietin receptor and decreases erythropoietin-stimulated cell proliferation. We show that Cis associates with the second tyrosine residue of the intracellular domain of the erythropoietin receptor (Tyr401). Two forms of Cis with molecular masses of 32 and 37 kDa were detected, and we demonstrate that the 37-kDa protein resulted from post-translational modifications of the 32-kDa form. Anti-ubiquitin antibodies recognized the 37-kDa form of Cis and the proteasome inhibitors N-acetyl-leucyl-leucyl-nor- leucinal and lactacystin inhibited its degradation, showing that the 37-kDa form of Cis is a ubiquitinated protein, which seems to be rapidly degraded by the proteasome. In erythropoietin-stimulated UT-7 cells, the activation of the erythropoietin receptor and signal transducer and activator of transcription 5 (STAT5) was transient and returned to basal levels after 30- 60 min of erythropoietin stimulation. In contrast, these proteins remained strongly phosphorylated, and STAT5 remained activated for at least 120 min in the presence of proteasome inhibitors. These experiments demonstrate that the proteasomes are involved in the down-regulation of the erythropoietin receptor activation signals. Because the proteasome inhibitors induced the accumulation of both the ubiquitinated form of Cis and the Cis- erythropoietin receptor complexes, our results suggest that the ubiquitinated form of Cis could be involved in the proteasome-mediated inactivation of the erythropoietin receptor.",
author = "Fr{\'e}d{\'e}rique Verdier and Stany Chr{\'e}tien and Odile Muller and Paule Varlet and Akihiko Yoshimura and Sylvie Gisselbrecht and Catherine Lacombe and Patrick Mayeux",
year = "1998",
month = "10",
day = "23",
doi = "10.1074/jbc.273.43.28185",
language = "English",
volume = "273",
pages = "28185--28190",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "43",

}

TY - JOUR

T1 - Proteasomes regulate erythropoietin receptor and signal transducer and activator of transcription 5 (STAT5) activation

T2 - Possible involvement of the ubiquitinated Cis protein

AU - Verdier, Frédérique

AU - Chrétien, Stany

AU - Muller, Odile

AU - Varlet, Paule

AU - Yoshimura, Akihiko

AU - Gisselbrecht, Sylvie

AU - Lacombe, Catherine

AU - Mayeux, Patrick

PY - 1998/10/23

Y1 - 1998/10/23

N2 - Cis is an Src homology 2 domain-containing protein, which binds to the erythropoietin receptor and decreases erythropoietin-stimulated cell proliferation. We show that Cis associates with the second tyrosine residue of the intracellular domain of the erythropoietin receptor (Tyr401). Two forms of Cis with molecular masses of 32 and 37 kDa were detected, and we demonstrate that the 37-kDa protein resulted from post-translational modifications of the 32-kDa form. Anti-ubiquitin antibodies recognized the 37-kDa form of Cis and the proteasome inhibitors N-acetyl-leucyl-leucyl-nor- leucinal and lactacystin inhibited its degradation, showing that the 37-kDa form of Cis is a ubiquitinated protein, which seems to be rapidly degraded by the proteasome. In erythropoietin-stimulated UT-7 cells, the activation of the erythropoietin receptor and signal transducer and activator of transcription 5 (STAT5) was transient and returned to basal levels after 30- 60 min of erythropoietin stimulation. In contrast, these proteins remained strongly phosphorylated, and STAT5 remained activated for at least 120 min in the presence of proteasome inhibitors. These experiments demonstrate that the proteasomes are involved in the down-regulation of the erythropoietin receptor activation signals. Because the proteasome inhibitors induced the accumulation of both the ubiquitinated form of Cis and the Cis- erythropoietin receptor complexes, our results suggest that the ubiquitinated form of Cis could be involved in the proteasome-mediated inactivation of the erythropoietin receptor.

AB - Cis is an Src homology 2 domain-containing protein, which binds to the erythropoietin receptor and decreases erythropoietin-stimulated cell proliferation. We show that Cis associates with the second tyrosine residue of the intracellular domain of the erythropoietin receptor (Tyr401). Two forms of Cis with molecular masses of 32 and 37 kDa were detected, and we demonstrate that the 37-kDa protein resulted from post-translational modifications of the 32-kDa form. Anti-ubiquitin antibodies recognized the 37-kDa form of Cis and the proteasome inhibitors N-acetyl-leucyl-leucyl-nor- leucinal and lactacystin inhibited its degradation, showing that the 37-kDa form of Cis is a ubiquitinated protein, which seems to be rapidly degraded by the proteasome. In erythropoietin-stimulated UT-7 cells, the activation of the erythropoietin receptor and signal transducer and activator of transcription 5 (STAT5) was transient and returned to basal levels after 30- 60 min of erythropoietin stimulation. In contrast, these proteins remained strongly phosphorylated, and STAT5 remained activated for at least 120 min in the presence of proteasome inhibitors. These experiments demonstrate that the proteasomes are involved in the down-regulation of the erythropoietin receptor activation signals. Because the proteasome inhibitors induced the accumulation of both the ubiquitinated form of Cis and the Cis- erythropoietin receptor complexes, our results suggest that the ubiquitinated form of Cis could be involved in the proteasome-mediated inactivation of the erythropoietin receptor.

UR - http://www.scopus.com/inward/record.url?scp=0032561322&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032561322&partnerID=8YFLogxK

U2 - 10.1074/jbc.273.43.28185

DO - 10.1074/jbc.273.43.28185

M3 - Article

VL - 273

SP - 28185

EP - 28190

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 43

ER -