Different Epidermal Growth Factor Receptor (EGFR) Agonists Produce Unique Signatures for the Recruitment of Downstream Signaling Proteins

Ronan, Tom; Macdonald-Obermann, Jennifer L.; Huelsmann, Lorel; Bessman, Nicholas J.; Naegle, Kristen M.; Pike, Linda J.

doi:10.1074/jbc.m115.710087

Cited by 53 publications

(58 citation statements)

References 43 publications

(14 reference statements)

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“…The sustained nature of the autophosphorylation signal at any ligand concentration suggests that it is indeed a feature of the EREG/EGFR or EPGN/EGFR complex rather than a consequence of low ligand affinity or receptor occupancy. Importantly, we also showed that amphiregulin promotes transient EGFR autophosphorylation (Figure S6D) despite binding the receptor with a similar (or lower) affinity than epiregulin or epigen (Ronan et al, 2016; Wilson et al, 2012). This is consistent with the ability of amphiregulin to induce sEGFR501 dimerization (Figure 4A), and further argues that reduced dimerization and sustained receptor activation are not simply consequences of low ligand/receptor affinity.…”

Section: Resultsmentioning

confidence: 68%

“…Numerous studies report distinct EGFR-dependent cellular responses to the different ligands (Wilson et al, 2009), with a given cell line responding differently to individual EGFR ligands in terms of cell proliferation (Wilson et al, 2012), differentiation (Kochupurakkal et al, 2005; Rizzi et al, 2013), and/or motility (Willmarth and Ethier, 2006). Individual EGFR ligands also induce qualitatively and quantitatively different downstream signals (Knudsen et al, 2014; Ronan et al, 2016; Wilson et al, 2012), and are linked to unique phenotypes in vivo (Wilson et al, 2009). …”

Section: Introductionmentioning

confidence: 99%

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EGFR Ligands Differentially Stabilize Receptor Dimers to Specify Signaling Kinetics

Freed

Bessman

Kiyatkin

et al. 2017

Cell

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Summary Epidermal growth factor receptor (EGFR) regulates many crucial cellular programs, with seven different activating ligands shaping cell signaling in distinct ways. Using crystallography and other approaches, we show how the EGFR ligands epiregulin (EREG) and epigen (EPGN) stabilize different dimeric conformations of the EGFR extracellular region. As a consequence, EREG or EPGN induce less stable EGFR dimers than EGF – making them partial agonists of EGFR dimerization. Unexpectedly, this weakened dimerization elicits more sustained EGFR signaling than seen with EGF, provoking responses in breast cancer cells associated with differentiation rather than proliferation. Our results reveal how responses to different EGFR ligands are defined by receptor dimerization strength and signaling dynamics. These findings have broad implications for understanding receptor tyrosine kinase (RTK) signaling specificity. Our results also suggest parallels between partial and/or biased agonism in RTKs and G protein-coupled receptors, as well as new therapeutic opportunities for correcting RTK signaling output.

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Section: Resultsmentioning

confidence: 68%

Section: Introductionmentioning

confidence: 99%

EGFR Ligands Differentially Stabilize Receptor Dimers to Specify Signaling Kinetics

Freed

Bessman

Kiyatkin

et al. 2017

Cell

Self Cite

298

347

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“…As mentioned above, most of the nodes in the signaling pathway followed the same direction in Estradiol, ICI 5 lg or ICI 5 mg groups compared with the control group. The direction of the genes corresponding to factors found within the cytoplasm could vary relative to the dynamics of the responses to the different ligands since it has been shown that recruitment of the downstream signaling proteins differs between EGFR agonists and also between different doses of the same agonist (Ronan et al 2016). However, it is interesting to note that the membrane receptor (EGFR) and one of the most important nuclear ligand-activated transcription factors for estradiol (ESR1) are upregulated in all the treatments, although the difference of expression for these genes between the treated and control fetuses in the general statistical analysis was not significant.…”

Section: Resultsmentioning

confidence: 99%

A transcriptomics model of estrogen action in the ovine fetal hypothalamus: evidence for estrogenic effects of ICI 182,780

2018

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Estradiol plays a critical role in stimulating the fetal hypothalamus–pituitary–adrenal axis at the end of gestation. Estradiol action is mediated through nuclear and membrane receptors that can be modulated by ICI 182,780, a pure antiestrogen compound. The objective of this study was to evaluate the transcriptomic profile of estradiol and ICI 182,780, testing the hypothesis that ICI 182,780 antagonizes the action of estradiol in the fetal hypothalamus. Chronically catheterized ovine fetuses were infused for 48 h with: vehicle (Control, n = 6), 17β‐estradiol 500 μg/kg/day (Estradiol, n = 4), ICI 182,780 5 μg/kg/day (ICI 5 μg, n = 4) and ICI 182,780 5 mg/kg/day (ICI 5 mg, n = 5). Fetal hypothalami were collected afterward, and gene expression was measured through microarray. Statistical analysis of transcriptomic data was performed with Bioconductor‐R and Cytoscape software. Unexpectedly, 35% and 15.5% of the upregulated differentially expressed genes (DEG) by Estradiol significantly overlapped (P < 0.05) with upregulated DEG by ICI 5 mg and ICI 5 μg, respectively. For the downregulated DEG, these percentages were 29.9% and 15.5%, respectively. There was almost no overlap for DEG following opposite directions between Estradiol and ICI ICI 5 mg or ICI 5 μg. Furthermore, most of the genes in the estrogen signaling pathway – after activation of the epidermal growth factor receptor – followed the same direction in Estradiol, ICI 5 μg or ICI 5 mg compared to Control. In conclusion, estradiol and ICI 182,780 have estrogenic genomic effects in the developing brain, suggesting the possibility that the major action of estradiol on the fetal hypothalamus involves another receptor system rather than estrogen receptors.

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“…The architecture of the EGFR signaling pathway is organized around a central repertoire of enzymes and adapter proteins that is capable of producing a large variety of cellular outcomes depending on the cellular context (1,14). The timing and extent of phosphorylation and effector recruitment varies, depending on which ligands and heterodimerization partners are present (60)(61)(62)(63). One explanation for the ability of different EGFR-family ligands to produce alternative phosphorylation and effector recruitment patterns is the intrinsic sequence specificity of EGFRfamily kinases, which can determine which sites become phosphorylated and able to support effector binding at different thresholds of kinase activity.…”

Section: Discussionmentioning

confidence: 99%

Deep mutational analysis reveals functional trade-offs in the sequences of EGFR autophosphorylation sites

Cantor

Shah

Kuriyan

2018

Preprint

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Upon activation, the epidermal growth factor receptor (EGFR) phosphorylates tyrosine residues in its cytoplasmic tail, which triggers the binding of Src Homology 2 (SH2) andPhosphotyrosine Binding (PTB) domains and initiates downstream signaling. The sequences flanking the tyrosine residues (referred to as phosphosites) must be compatible with phosphorylation by the EGFR kinase domain and the recruitment of adapter proteins, while minimizing phosphorylation that would reduce the fidelity of signal transmission. In order to understand how phosphosite sequences encode these functions within a small set of residues, we carried out high-throughput mutational analysis of three phosphosite sequences in the EGFR tail.We used bacterial surface-display of peptides, coupled with deep sequencing, to monitor phosphorylation efficiency and the binding of the SH2 and PTB domains of the adapter proteins Grb2 and Shc1, respectively. We found that the sequences of phosphosites in the EGFR tail are restricted to a subset of the range of sequences that can be phosphorylated efficiently by EGFR.Although efficient phosphorylation by EGFR can occur with either acidic or large hydrophobic residues at the −1 position with respect to the tyrosine, hydrophobic residues are generally excluded from this position in tail sequences. The mutational data suggest that this restriction results in weaker binding to adapter proteins, but also disfavors phosphorylation by the cytoplasmic tyrosine kinases c-Src and c-Abl. Our results show how EGFR-family phosphosites achieve a trade-off between minimizing off-pathway phosphorylation while maintaining the ability to recruit the diverse complement of effectors required for downstream pathway activation.Cantor et al.

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Different Epidermal Growth Factor Receptor (EGFR) Agonists Produce Unique Signatures for the Recruitment of Downstream Signaling Proteins

Cited by 53 publications

References 43 publications

EGFR Ligands Differentially Stabilize Receptor Dimers to Specify Signaling Kinetics

EGFR Ligands Differentially Stabilize Receptor Dimers to Specify Signaling Kinetics

A transcriptomics model of estrogen action in the ovine fetal hypothalamus: evidence for estrogenic effects of ICI 182,780

Deep mutational analysis reveals functional trade-offs in the sequences of EGFR autophosphorylation sites

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