Identification of both positive and negative domains within the epidermal growth factor receptor COOH-terminal region for signal transducer and activator of transcription (STAT) activation

Ling Xia, Lijuan Wang, Alicia S. Chung, Stanimir S. Ivanov, Mike Y. Ling, Ana M. Dragoi, Adam Platt, Tona M. Gilmer, Xin Yuan Fu, Y. Eugene Chin

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

The cytoplasmic region of human epidermal growth factor receptor (EGFR) contains an intrinsic tyrosine kinase (697-955) followed by a 231-residue-long COOH-terminal tail (C-tail), which contains multiple tyrosine residues. To examine the role of the EGFR C-tail in signal transducer and activator of transcription (STAT) activation, a series of EGFR C-tail truncations were constructed. Transient transfection of 293 cells with EGFR lacking the C-tail, i.e. Y974ΔEGFR or Y992ΔEGFR, led to EGF-independent or constitutive STAT activation, whereas EGF-dependent STAT activation was restored with truncations made COOH-terminal to the next tyrosine residue, i.e. EGFR-Y1045Δ. Transfection with the truncated form EGFR-Y954Δ resulted in the loss of STAT activation, suggesting that the sequence between Tyr 974 and Tyr 954 is essential for STAT activation. Phosphopeptide competition analysis revealed multiple tyrosine residues within the C-tail that can act as the docking sites for both Stat1 and Stat3. A region that negatively regulated STAT activation was also identified, extending from Tyr 1114 to Glu 1172, consistent with the ability of this region to recruit a suppressor of cytokine signaling factors SOCS1 and SOCS3. When cotransfected with the full-length EGFR, but not Y992ΔEGFR, SOCS1 or SOCS3 inhibited STAT activation by EGF in 293 cells. This suggests that both SOCS1 and SOCS3 can negatively regulate EGFR activation, presumably by inducing ubiquitination-dependent EGFR degradation upon ligand binding. These findings may therefore offer clues to how the EGF receptor C-tail regulates STAT activity.

Original languageEnglish (US)
Pages (from-to)30716-30723
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number34
DOIs
StatePublished - Aug 23 2002
Externally publishedYes

Fingerprint

Transcription
Transducers
Epidermal Growth Factor Receptor
Transcriptional Activation
Chemical activation
Tail
Epidermal Growth Factor
Tyrosine
Transfection
Phosphopeptides
Ubiquitination
Protein-Tyrosine Kinases
Cytokines
Ligands
Degradation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of both positive and negative domains within the epidermal growth factor receptor COOH-terminal region for signal transducer and activator of transcription (STAT) activation. / Xia, Ling; Wang, Lijuan; Chung, Alicia S.; Ivanov, Stanimir S.; Ling, Mike Y.; Dragoi, Ana M.; Platt, Adam; Gilmer, Tona M.; Fu, Xin Yuan; Eugene Chin, Y.

In: Journal of Biological Chemistry, Vol. 277, No. 34, 23.08.2002, p. 30716-30723.

Research output: Contribution to journalArticle

Xia, Ling ; Wang, Lijuan ; Chung, Alicia S. ; Ivanov, Stanimir S. ; Ling, Mike Y. ; Dragoi, Ana M. ; Platt, Adam ; Gilmer, Tona M. ; Fu, Xin Yuan ; Eugene Chin, Y. / Identification of both positive and negative domains within the epidermal growth factor receptor COOH-terminal region for signal transducer and activator of transcription (STAT) activation. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 34. pp. 30716-30723.
@article{acf39a6881114daa80fb8925f4400b30,
title = "Identification of both positive and negative domains within the epidermal growth factor receptor COOH-terminal region for signal transducer and activator of transcription (STAT) activation",
abstract = "The cytoplasmic region of human epidermal growth factor receptor (EGFR) contains an intrinsic tyrosine kinase (697-955) followed by a 231-residue-long COOH-terminal tail (C-tail), which contains multiple tyrosine residues. To examine the role of the EGFR C-tail in signal transducer and activator of transcription (STAT) activation, a series of EGFR C-tail truncations were constructed. Transient transfection of 293 cells with EGFR lacking the C-tail, i.e. Y974ΔEGFR or Y992ΔEGFR, led to EGF-independent or constitutive STAT activation, whereas EGF-dependent STAT activation was restored with truncations made COOH-terminal to the next tyrosine residue, i.e. EGFR-Y1045Δ. Transfection with the truncated form EGFR-Y954Δ resulted in the loss of STAT activation, suggesting that the sequence between Tyr 974 and Tyr 954 is essential for STAT activation. Phosphopeptide competition analysis revealed multiple tyrosine residues within the C-tail that can act as the docking sites for both Stat1 and Stat3. A region that negatively regulated STAT activation was also identified, extending from Tyr 1114 to Glu 1172, consistent with the ability of this region to recruit a suppressor of cytokine signaling factors SOCS1 and SOCS3. When cotransfected with the full-length EGFR, but not Y992ΔEGFR, SOCS1 or SOCS3 inhibited STAT activation by EGF in 293 cells. This suggests that both SOCS1 and SOCS3 can negatively regulate EGFR activation, presumably by inducing ubiquitination-dependent EGFR degradation upon ligand binding. These findings may therefore offer clues to how the EGF receptor C-tail regulates STAT activity.",
author = "Ling Xia and Lijuan Wang and Chung, {Alicia S.} and Ivanov, {Stanimir S.} and Ling, {Mike Y.} and Dragoi, {Ana M.} and Adam Platt and Gilmer, {Tona M.} and Fu, {Xin Yuan} and {Eugene Chin}, Y.",
year = "2002",
month = "8",
day = "23",
doi = "10.1074/jbc.M202823200",
language = "English (US)",
volume = "277",
pages = "30716--30723",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "34",

}

TY - JOUR

T1 - Identification of both positive and negative domains within the epidermal growth factor receptor COOH-terminal region for signal transducer and activator of transcription (STAT) activation

AU - Xia, Ling

AU - Wang, Lijuan

AU - Chung, Alicia S.

AU - Ivanov, Stanimir S.

AU - Ling, Mike Y.

AU - Dragoi, Ana M.

AU - Platt, Adam

AU - Gilmer, Tona M.

AU - Fu, Xin Yuan

AU - Eugene Chin, Y.

PY - 2002/8/23

Y1 - 2002/8/23

N2 - The cytoplasmic region of human epidermal growth factor receptor (EGFR) contains an intrinsic tyrosine kinase (697-955) followed by a 231-residue-long COOH-terminal tail (C-tail), which contains multiple tyrosine residues. To examine the role of the EGFR C-tail in signal transducer and activator of transcription (STAT) activation, a series of EGFR C-tail truncations were constructed. Transient transfection of 293 cells with EGFR lacking the C-tail, i.e. Y974ΔEGFR or Y992ΔEGFR, led to EGF-independent or constitutive STAT activation, whereas EGF-dependent STAT activation was restored with truncations made COOH-terminal to the next tyrosine residue, i.e. EGFR-Y1045Δ. Transfection with the truncated form EGFR-Y954Δ resulted in the loss of STAT activation, suggesting that the sequence between Tyr 974 and Tyr 954 is essential for STAT activation. Phosphopeptide competition analysis revealed multiple tyrosine residues within the C-tail that can act as the docking sites for both Stat1 and Stat3. A region that negatively regulated STAT activation was also identified, extending from Tyr 1114 to Glu 1172, consistent with the ability of this region to recruit a suppressor of cytokine signaling factors SOCS1 and SOCS3. When cotransfected with the full-length EGFR, but not Y992ΔEGFR, SOCS1 or SOCS3 inhibited STAT activation by EGF in 293 cells. This suggests that both SOCS1 and SOCS3 can negatively regulate EGFR activation, presumably by inducing ubiquitination-dependent EGFR degradation upon ligand binding. These findings may therefore offer clues to how the EGF receptor C-tail regulates STAT activity.

AB - The cytoplasmic region of human epidermal growth factor receptor (EGFR) contains an intrinsic tyrosine kinase (697-955) followed by a 231-residue-long COOH-terminal tail (C-tail), which contains multiple tyrosine residues. To examine the role of the EGFR C-tail in signal transducer and activator of transcription (STAT) activation, a series of EGFR C-tail truncations were constructed. Transient transfection of 293 cells with EGFR lacking the C-tail, i.e. Y974ΔEGFR or Y992ΔEGFR, led to EGF-independent or constitutive STAT activation, whereas EGF-dependent STAT activation was restored with truncations made COOH-terminal to the next tyrosine residue, i.e. EGFR-Y1045Δ. Transfection with the truncated form EGFR-Y954Δ resulted in the loss of STAT activation, suggesting that the sequence between Tyr 974 and Tyr 954 is essential for STAT activation. Phosphopeptide competition analysis revealed multiple tyrosine residues within the C-tail that can act as the docking sites for both Stat1 and Stat3. A region that negatively regulated STAT activation was also identified, extending from Tyr 1114 to Glu 1172, consistent with the ability of this region to recruit a suppressor of cytokine signaling factors SOCS1 and SOCS3. When cotransfected with the full-length EGFR, but not Y992ΔEGFR, SOCS1 or SOCS3 inhibited STAT activation by EGF in 293 cells. This suggests that both SOCS1 and SOCS3 can negatively regulate EGFR activation, presumably by inducing ubiquitination-dependent EGFR degradation upon ligand binding. These findings may therefore offer clues to how the EGF receptor C-tail regulates STAT activity.

UR - http://www.scopus.com/inward/record.url?scp=0037163125&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037163125&partnerID=8YFLogxK

U2 - 10.1074/jbc.M202823200

DO - 10.1074/jbc.M202823200

M3 - Article

VL - 277

SP - 30716

EP - 30723

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 34

ER -